Figures and Figure Legends

This is part of a series of tutorials I’m putting together for my students.

So, you have collected some interesting data from your experiments. Since no one but you will be reading your lab notebook (but hopefully people could if they wanted to), you need to present that data in figures so the rest of the world can know what you did and decipher your results.

Deciding what data gets to be a manuscript figure

The purpose of your manuscript is to show evidence for a new conclusion and the data presented in your figures should tell this story. Remember, the order of your figures for your manuscript may not necessarily (and probably won’t be) the order in which you collected the data. So, once you have all of your figures assembled, print them out one per page and work on defining the best order of presentation to make the case for your new conclusion. Now is a good time to evaluate whether there are any potential weaknesses regarding support for your conclusions, either in the data you already have or data you may still need to collect. You want to present the strongest possible case before your manuscript is submitted for review, but everything is a cost-benefit analysis and you’re always against the clock.

At this point you may also notice that some of the data presented in figures may be tangential, not quite fit with the rest or break up the flow of the story you are trying to tell. Authors can decide to cull certain figures (Sorry, data you have to remain in the lab notebook) or move them to ‘Supplementary Figures.’ Many journals allow for the inclusion of Supplementary Material- extra figures, longer versions of methods, large tables of data or files that would never be appropriate for print format. These Supplementary Materials exist as electronic files only linked to your final accepted manuscript as it appears as a journal article. Each journal has a different policy for what is acceptable for Supplementary Material. Some are more inclusive- the more data the merrier, drag everything out of all authors’ lab notebooks. Others are very limiting- essential data and files in appropriate for print only and anything else must be incorporated into the main body of the manuscript or cut out completely.

Preparing figures starts with high-quality data.

Images should be of sufficient resolution. Any adjustments of brightness and contrast must be made to the entire image; adjusting selective portions is unethical data manipulation and scientific fraud. Cropping is OK, but again beware of excessive image manipulations. They are usually an indicator that you need to repeat the experiment to obtain the necessary data.

Experimental data should be free of technical errors or other artifacts. The results should come from experiments as described in the methods section. Consistency in following experimental protocols (and including all of those details in your notebook) should be standard lab practices. Controls must be performed for each experiment so that the results can be properly interpreted. As you evaluate the figures you have made from your data, check again to see if all necessary controls have been included. When in doubt, don’t skimp on this- repeat the experiment with the proper controls. Your co-authors and reviewers will likely eventually tell you the same thing.

Your data should also be repeated enough times to be statistically relevant. Note that this does not mean you repeat an experiment enough times until you get the data you want. This ‘cherry-picking’ is another unethical manipulation of data. Unfortunately, this type of fraud is the most difficult to catch by the peer-review system. Reviewers have no way of knowing that you have a hundred other experimental trials with contradictory data in your lab notebook. Scientists must have the integrity to accurately present their results and have legitimate justifications for excluding some data (altered variables, confounding variables, improper controls etc). It is not always possible to show all repetitions of an experiment and in some cases (like gels) it is not even feasible to average the results. Showing ‘representative data’ (a single instance of the most common result) is perfectly acceptable, but it should be just that- representative of your average results.

Don’t pursue perfect data at the expense of integrity. The rising standards of scientific work and competition for rewards based on that work create an enormous amount of pressure to compromise your integrity for the sake of publication. RESIST! Research fraud undermines our entire enterprise. Biological systems are inherently complex and imperfect- we should not expect results to be simple and pristine.  Control for what you can when you can, but do not otherwise force data to yield a certain result.

Putting together figure files

Usually your data will consist of images or graphs. These electronic files must be edited to include the raw data as well as appropriate labels. The simplest way of doing this is to drop the images into a PowerPoint slide to assemble all the necessary parts. Text boxes can be used to add labels. Lines and arrows can be added to draw attention to certain features. All labels and features of your figures should be properly aligned using the automatic tools for doing so. More complex figures consist of multiple parts that are designated by letters (Ex: Figure 1A and Figure 1B), and these letters can be added as text boxes. Journals tend to have preferences for the exact labeling details (fonts, sizes etc) and the instructions to authors will have this information. Make sure you read this information carefully and apply it consistently across all figures. Don’t use Arial capital letters to label the parts of Figure 1, Calibri Roman numerals for Figure 3 and lower case Times New Roman on Figure 5. You’re not in cloud cuckoo land. Editors, reviewers and other scientists appreciate consistency.

Remember that in the final manuscript format, the sizes of all labels and images will be considerably reduced. Make sure that your figure as submitted in manuscript form is sufficiently large so that it is still interpretable at a much smaller size. Any lines on graphs should be of sufficient thickness so as not to disappear or lose their pattern upon reduction. Note that it is generally easier to number samples like gel lanes, mulitpart images, etc than to write out the full sample description in the figure. Save the full sample names and descriptions for the figure legend.

When available, move up in the food chain to a program like Adobe Photoshop or Illustrator or Corel Draw to put together figure images. These programs have a steeper learning curve, but offer more sophisticated options for putting the figure file together and saving it as a high resolution image. For many journals, your figures must be submitted as image files (usually .tiff) or as PDF pages. Most journals use the manuscript submission phase as their quality control phase, meaning the files you submit for review must be of sufficient quality for the manuscript proof. Speaking of higher quality software, programs like OriginLab and Kaleidagraph are much better at generating image quality graphs than Microsoft Excel.

Color vs. Black/White or Grayscale Figures

Journals will typically charge you more to print color figures over black and white or grayscale images. (Oh, so yeah, if you didn’t get the memo, the authors typically pay publication charges to cover the printing and/or access for the published work. But then again, if you’ve gotten this far, you’ve realized you’re not in science for the money.) When possible use black and white or grayscale figures. If graphs become too complicated in monotone, try breaking up the number of samples shown on the same axis. Of course, you shouldn’t completely eschew color. Use it when it is most appropriate to distinguish samples. For example, it’s not that big of a deal to show a Coomassie-stained gel in black and white, but pictures of Arabidopsis showing wild-type and mutant plants with varying degrees of pigmentation should definitely be in color. Finally, as part of the ‘use color judiciously rule’, stick with the basic colors (8 crayola box, not the 196) so that there are no incompatibilities or unexpected shifts in tone when transferring files or changing file-types. Also keep in mind that there is ~10% prevalence of red-green colorblindness, so avoid using these colors together to differentiate between key samples. (Hey, after #TheDress this week, maybe you should just avoid color altogether.)

The figure heading, title and legend

Each figure should have a heading as defined by the journal (Ex: ‘Figure 1.’ ‘Fig. 1’ or ‘Figure 1:’). Each figure should also have a title, the formatting of which may be explicitly defined by the journal. It may be required to be in the form of a complete sentence or just a concise phrase; it may be required to be in bold or italics to distinguish it from the legend. The legend should tell the reader what they are looking at. It is not necessary to include lengthy procedural details, but it is useful to mention the name of the experiment and any details about treatment or sample preparation useful for interpreting the data in the figure. It should define all parts shown. Every sample or label on the figure must be defined in the legend.

Other random and lesser commandments

  1. Thou shalt be consistent across all figures.
  2. Thou shalt not use yellow for graph lines.
  3. Thou shalt include error bars.
  4. Thou shalt have elements sized appropriately relative to one another.

Include your figure and figure legend tips and lesser commandments below.

Johnna

Literature Searching, Reading, Organizing

This is part of a tutorial series I’m writing for my students. Science writing first requires a lot of science reading.

Any scientific manuscript will require numerous other references to scientific literature to substantiate the facts upon which it builds. This means you have to become familiar with a body of literature related to the topic. Finding reliable references and sorting out what they mean is no small task. As a scientist, it is useful to make literature searching and reading a regular part of your routine. Set a goal to read a certain number of papers each week to keep up with the research in your area. When you are in ‘writing-mode’ for a grant or a scientific manuscript, the reading will likely be more intense, but it is a general good practice to keep up with the scientific literature a little bit at a time.

Searching for Literature

My go-to search engines for finding scientific literature are Google Scholar and PubMed. You can search key words, titles, authors, year, narrow by article type (review, patent, clinical study, research article etc). There are also numerous options for narrowing down your search and sorting the results (relevance, dates etc). The links for the titles can generally take you to the full text of the article (beware of paywalls if you’re not on a network with an institutional subscription). If based on the abstract of the article, you’d like to get a look at the full text, but you’re on the wrong side of a paywall, you could always e-mail the corresponding author to request a copy. For a quicker response, tweet the reference with the hashtag #ICanHazPDF with email [at] domain.com and someone out there on the interwebz with access will send it to you.  One of the biggest issues you will likely have is sifting through the long list of titles for something that is actually useful to you. When embarking on a new literature search, try to find a recent review article to give you an overview of the topic and point you to relevant primary research articles. Then just start reading and following citations through the literature until you have all of the information you need.

https://scholar.google.com/

http://www.ncbi.nlm.nih.gov/pubmed

 

Reading Primary Research Literature

The best way to understand scientific papers is to practice reading them. There’s a learning curve for the jargon and background in your field, but sometimes papers are also just crappily written. By nature, scientific literature is information dense and since it continually builds on previous studies, the reader always enters in the middle of the action. If the paper is well-written, the abstract should give you a sense of the importance of the work, the research that was done and what it means. The introduction should give you just enough information to allow you to understand the research question that will be addressed. In primary research papers, the introductions should be fairly focused. Consult a review article for a broader scope of what’s going on in any given research area. Unless you are looking for a particular protocol or are trying to replicate an experiment, the methods section probably won’t get much attention reading through an article for the first time. The first time through, just get an idea of the techniques used in the research, but wait to sort out any of the finer details until you see the figures in the results section. As you’re sorting through the results section, this is where the authors are showing you their data. They should explain some of the rationale behind the line of experiments, what data they collected and what it means. There should be some connection or flow among the figures and results that ultimately builds up to (an) overall conclusion(s). In some articles, the data are presented to build a case for a certain model or overall conclusion. In others, the experiments are geared towards eliminating possibilities until the results focus in on a particular conclusion. Many papers aren’t always written this way and there is a greater burden on the reader to interpret the results to see if it leads you to the same conclusion. Personally, I like papers that can end with some sort of model (cartoons are appreciated) to visually sum up all of the conclusions. In any case, the reader should give a critical eye to all results in the relation to how they are supporting the conclusions. Look at the figures and write down your own results; are they the same as the description the authors give? Ask yourself if the results mean what the authors say they mean. Could there be any other possible interpretation of the results? Then think deeper about the data they are showing. Are those experiments the best way of figuring out what’s going on? Is the data of sufficient quality (error bars, statistics, clarity in images etc)? Did they perform all of the appropriate controls? Refer back to the methods section for finer points of the protocols. Are there any red flags about how the work was performed that could influence the results? The peer-review process is not perfect, so even though it is science’s way of validating work prior to publication, it doesn’t mean that there are no mistakes or misinterpretations. After all, the science was performed, written and evaluated by humans.

Keeping up with the Literature

There are a few ways to make sure you are keeping up with the literature in your field of interest. Set aside some time each day or week to at least scan the titles of what’s going on in your research area. Set a goal for paper-reading, even starting out at 1 paper/week will be useful. Check out the hashtag #365papers on Twitter for inspiration and accountability. There are a few different ways to automate the process as well. Sign up for eTOC (electronic table of contents) alerts from journals you read often. The contents of each issue will be delivered to your inbox as soon as they are available. Give them a quick scan for interesting key words and download what seems interesting or relevant. Harness the power of the Google search engine and sign up for Google Scholar alerts. Based on keywords that you provide, Google Scholar will send you daily updates with relevant literature.

Organizing Literature

As a scientist, you will begin to amass numerous downloaded PDF files of research articles. It’s your own digital library for the full-text version of useful papers. For many, this collection is purely electronic. Gone are the days of file cabinets filled to the brim with reprints and papers with scribbled notations in the margins. If you prefer, printing and hand-writing notes is still possible, but not feasible for every paper you will ever read. So, you need to come up with an organization system that works for you. I have a folder where I dump all of my papers and they all have filenames (year first author name). It’s rudimentary, but it works for me. I can often remember which paper I need to open, but when I can’t it’s an easy search. Writing this tutorial has shown me that I’m still in the dark ages with respect to my organization strategy. There are a number of software options for helping you manage your own digital library. Check out Papers, Mendely, CiteULike. These programs offer more options for searching and sorting your own digital library as well as some integration with internet-wide literature searching.

Give your favorite searching, reading and organization tips in the comments below. Please weigh in on a literature organization/management software preference because I’m embarrassed to have not gotten with one of these programs yet. #OldSchool #AtLeastItsNotPaper

Johnna

Writing the Materials and Methods Sections

This is part of a tutorial series I’m writing for my students on scientific manuscript preparation.

This section should include information on every experimental procedure used in the work. You don’t have to include experimental details of everything that was in your lab notebook or all of the optimization work you did for the project. Yeah, readers don’t care about the two-years’ worth of failed protein purification methods in your notebook. You will only include the short paragraph of what did finally work. Refer to your figures. Whatever experiments yielded data that is ultimately found in your figures should have details in your Materials and Methods section. If certain reagents were critical to completion of the experiments, then include your sources for them (commercial or donations from other labs). The Materials and Methods section is written in the past tense without the use of personal pronouns. It may seem awkward to get into this point of view, but there is an omniscient perspective from which your methods can be written. I’m not sure where this convention originated, but remove yourself from the experiment and it seems more legitimate.

Subdivide this section into useful parts. This kind of organization also helps the reader pinpoint details for replication or evaluation of experiments. Often readers will be looking at your work to perform similar experiments or repeat your work.

It’s not exactly a protocol, but it should be complete. You don’t have to tell your readers every step of a protocol, but it should be complete enough for someone else with comparable training to replicate. The literature varies widely in the usefulness and clarity of methods descriptions. For more complex procedures, authors are often contacted directly for lab protocols or additional information not included in the materials and methods section. As methods have become more complex, fewer details are included. This is especially true of molecular biology work. Typically primer sequences are given and specific vectors are mentioned, but it can be especially irksome to figure out others’ cloning construction strategies. Usually, though, this information can be obtained directly from the authors. In general, I would err on the side of completeness (vs. concise) because this makes it easier for your experimental strategies to be evaluated and replicated. This strategy can run afoul of the journal’s character limits, but in this age of online publication, supplemental information is allowed. You can write a ‘Supplemental Methods’ section which will be available online only and can be as long as you’d like.

Referencing previous literature is a convenient way of being concise. It is acceptable to say something like, “This experiment was performed as in Reference (1).” However, if you go this route, make sure that the reference you are citing in this case does have the necessary information on experimental details. The scientific literature is infamous for pulling this trick only to reference yet another paper that says “This experiment was performed as in Reference (2).” You may have to follow this rabbit hole back a couple of decades before you find any real useful information on performing the experiment. (This is a personal pet peeve of mine that just makes me feel stabby when I encounter this nonsense.) So save your readers some trouble and either include the necessary info or cite a direct source. For many labs, there are standard references and descriptions for routinely used methods that appear in almost all of the manuscripts. It’s fine to revert back to previously published articles to see how things were worded and what references were cited, but it’s always a good idea to verify these references yourself. Otherwise, a mis-referenced citation will be propagated many times over. (This is also a good practice for any citation anywhere in a manuscript- to make sure the reference is really saying what you think it does.)

Offer your pro-tips for methods-writing in the comments section below.

Johnna

Citations and Reference Formatting in Manuscripts

This is the next post in a series of tutorials I’m writing for my students related to scientific manuscripts.

For every statement of fact in your introduction, results or conclusions sections or for every method, it is important to cite the appropriate reference, which provides the basis for this fact or method resource. Thus, references are given throughout the manuscript to direct the reader to previous literature to independently verify the facts or otherwise track down more information. Providing references for information in a scientific manuscript is somewhat different than providing references in a research paper for the humanities. While there may be some similarities with MLA style, there are specific requirements for scientific manuscripts that are determined by the choice of journal for submission.

Acceptable references are books and other peer-reviewed articles. You’re not allowed to get facts from random sites on the internet and other unreliable sources. Scientific literature is vetted by a peer-review process, in which other scientists evaluate the merit of submitted work. While it’s not perfect, this arduous process of evaluation and revision is science’s way of validating reports of experimental work. It means you can trust the information within peer-reviewed journal articles more than you can the random blog post or Wikipedia entry.

Specific formatting for references can be found in the instructions for authors. Some journals prefer using numbers in parentheses, some use superscripts, and some use an (author, year) formatting within the text of the manuscript. Reference lists are likewise variable according to journal- numbered lists, alphabetical lists, include titles, no titles, full journal names, abbreviated journal names etc. etc. In any case, the format will include enough basic information for a reader to look up the exact reference and check your facts. You have to follow the exact style for the journal of choice or the editor in charge of looking over new manuscript submissions will notice and kick your manuscript back to you- not reviewed, not accepted, not any further beyond your lab notebook than your computer files.

Citation management software should be your new best friend. Chances are the previous written assignments you’ve had haven’t required dozens of citations backing up various facts in your manuscript. You also probably worked on all of your previous manuscripts from start to finish in the order in which they will appear. As stated in my previous tutorial, you generally work on the sections of your scientific manuscript in a very different order than the one in which it will be finally assembled. Not to mention that working with multiple authors means merging sections from different writers and major rearrangements of written work. All of these complications mean that it is futile to try to manually keep track of all of the references that go along with these edits.This is a waste of your time and will lead to errors. Use citation management software. Citation management software like EndNote allows you to search for literature, manage all relevant information for a citation and integrate it into your handy text editing software (MS Word) in a way that lets you point-and-click which references to incorporate. This software will re-order citations in the reference list as sections are rearranged. The citation management software also already knows the exact details of each journal’s citation style. Simply select the appropriate journal option in the citation management software and EndNote will keep track of journal abbreviations, what should be italicized, what things are separated by commas and where periods should go. And if you decide to switch the journal for submission, all you have to do is click a different button and at least the reference section is done. Using citation management software ensures that no random numbers in parentheses are forgotten in the middle of your text.

Here’s a brief tutorial for using EndNote.

The EndNote software allows you to create a library of citations that can be used in your writing projects. There are two general strategies for EndNote library usage. (1) A single EndNote library containing every citation you will ever use. (2) Multiple EndNote libraries with one for each writing project. Each has its own merits and drawbacks. I know people that use both successfully, so if you feel strongly one way or the other, go with the one that seems most intuitive to you.

Citations can be added to your EndNote reference library by using the on-line search function. The most useful searches for scientific manuscripts are the PubMed (NLM) and Web of Science (ISI) searches. If you enter an author, publication year, journal name and some relevant keyword, EndNote should easily identify the necessary reference, which can then be saved in your library. It should be noted that I find this search feature useful for finding specific references (like those you’ve already read and know you want to use) rather than a general search to keep up with the literature (then again, I’m a type 2 EndNote user).

Reference entries can be added manually to an EndNote library also. If the paper is ancient or otherwise impossible to find with the search tools, you can manually type in all of the fields.

Duplicate entries within a library can be identified and eliminated. This should be done or it can cause problems when trying to use these entries while writing in MS Word.

The EndNote program also integrates itself as an add-on feature in MS Word. Once installed, it will have its own tab in the top toolbar. While working on a manuscript in MS Word, open the EndNote program and the appropriate library. From the EndNote tab in MS Word, all you have to do is click the ‘Insert Citation’ button to select the appropriate reference from the library. Note, the reference must be a part of the library at this point (already captured from an on-line search) because you can’t do an on-line search via EndNote from MS Word. Once selected, a reference placeholder will appear in the text (whether (#) or (author, date)) and a reference list will be started at the end of the text portion of your MS Word file. All of the details are determined by the designated style, given in terms of the journal title, which can also be changed from a drop-down style menu in the EndNote tab in MS Word.

Finer points of citation formatting with EndNote…

You can make some changes to the style if it’s not exactly correct based on the most up to date info on the journal’s website.

If you are citing multiple references at the end of a particular sentence, you can reorder them according to the stipulations of the journal style (usually in chronological order, oldest to most recent).

An EndNote library can be exported from an MS Word file. So, if you get a written portion from another author, you can recreate their EndNote library of citations even if they didn’t send you their library file.

A note on plagiarism: Don’t do it. Just because you are citing a reference in order to substantiate a statement in your manuscript doesn’t mean that you can copy their exact words. Use your own words and incorporate their work as it relates to yours. Don’t sweat whether minor wording changes are enough. Since you are building on these previous facts, the context will be different than the original paper, but there’s no excuse for copying things word for word. Sure, there are limited numbers of ways of saying any given truth clearly and concisely, but you can find a way of stating a necessary fact in your own words. Plus, you should really understand the previous literature well enough to restate necessary information in the context of your research manuscript without stealing someone else’s words.

Share your reference, citation and EndNote pro-tips in the comments section below.

Johnna

Clear and Concise: General Suggestions for Manuscript Preparation

Today’s post is the first tutorial on some general good practices of scientific writing and manuscript preparation. Many of the suggestions are based on common errors I’ve seen so far in very rough drafts from my students. If you have other suggestions, please offer them in the comments section.

Congratulations, you have some research to report to the rest of the world. No one knows it better than you, and it’s your job to educate the rest of us. The whole point is to clearly convey what you did, what it means and why it’s important.

It may seem overwhelming where to begin, but the specifics of sections and formatting have largely been decided for you depending on the journal you have chosen* for your manuscript submission. Each scientific journal provides detailed ‘instructions to authors’ on their websites. It includes everything you need to know about putting your manuscript document(s) together- section headings, font and size, citation formatting and figure guidelines. Some even provide a useful template document for MS Word. It is imperative that you follow these guidelines explicitly (yes, all of them. yes, exactly) or your research manuscript will be bounced back to you by the editor without review until you make it conform to their requirements. If you have any questions about how your document should look, download an article from that journal and/or have a look at an available pre-publication accepted manuscript version; each readily available at the website for that journal.

Outside of the requirements of the journal, it’s up to your good practice and judgment to put together a written document that clearly conveys your research. There are typically character or page limitations so it is important to be concise. State your thoughts in the most direct and clear way. This style may not win you a Pulitzer prize, but your audience is reading for information content, not necessarily pleasure. Verbose descriptions or filler sentences that don’t really say much can be a frustrating waste of time for the reader.  At the other end of the spectrum, it shouldn’t be so terse as to be incomprehensible due to missing information or no connection with anything of greater significance. (It should be noted that your manuscript’s first readers will be an editor and reviewers that will decide whether your research ever sees the light of day beyond your lab notebook and documents on your computer. So, yeah, at the very least- it shouldn’t be difficult or frustrating to read.)

Another good piece of general advice is ‘be consistent.’ Consistency- in terms of abbreviation usage and nomenclature in the text and presentation in the figures. If you refer to an organism one way in one paragraph, it should be same in subsequent paragraphs, unless you are talking about a different organism. While synonyms can be useful to break up the monotony, for important things like technique names and critical variables, refer to them in the exact same way or the reader may think you’re talking about something else. It is also useful to consistently order/label samples in your figures. If untreated sample is a solid black line and treated sample is a dashed line in Panel A, then don’t switch them for Panel B. This consistency should propagate across figures as well. If wild type is to the left of the mutant in images or lanes on a gel for a figure, then don’t switch the orientation in subsequent figures.

Abbreviations- you should use them. For any uncommon and long gene/protein names or techniques names, use an abbreviation. Write out the full-length version of the name or technique at its first usage and follow it by the abbreviation in parentheses. Then, just use that abbreviation in the rest of the manuscript. The general rule is, if you’ll need to use cumbersome wording 3 times or more, use an abbreviation. There are a number of very commonly used abbreviations within a scientific field that are widely understood and do not need definitions (ex: DNA, RNA, ATP etc) and these are generally listed somewhere on the website for each journal. Manuscripts also have an abbreviations section somewhere at the very beginning or end where all abbreviations are defined in one place. If the reader missed the definition in the text, they can refer to the abbreviations section. Abbreviations should be used consistently and exactly as defined. (Ex: Photosystem II = PS II; not sometimes PSII or PsII etc).

Specific usage and nomenclature- organisms, genes, proteins. This nomenclature often gets mixed in biochemistry literature. The full proper names of any organisms should be given the first time they are mentioned with the genus name capitalized and italicized and the species name lowercase and italicized. (Ex: Drosophila melanogaster, Zea mays, Arabidopsis thaliana) Then, if you plan on referring to this organism other places throughout the manuscript, designate the shortened version in parentheses after the first usage. (Ex: …. Drosophila melanogaster (subsequently Drosophila)….) Just make sure you use exactly that shortened designation consistently throughout the rest of the manuscript because any deviation from that usage will make the reader wonder if you are referring to another species. The specific shortened designations can vary widely, some people just stick with Genus others G. species and for some reason some organisms are so commonly known by their genus name that it has become acceptable to write Genus (nonitalicized) like Arabidopsis. There are strict conventions for the proper designations of gene or protein names/abbreviations. In a more genetics/cell biology manuscript, the rules are stricter**, but in more biochemically-focus papers, we’re not such sticklers. That being said, again, consistency is key and it should be clear whether you are talking about a protein or a gene. Typically genes are italicized (Ex: adh, Adh, ADH) and proteins are nonitalicized and at least the first letter is capitalized (Ex: Adh, ADH).

Scientific writing is generally written in third person format without any personal pronouns. It may take some getting used to this style, but eventually you will find ways of writing methods and results without any mention of yourself. (Ex: The colonies were screened using PCR. vs. We isolated DNA and did PCR.) Also, the Methods and Results are typically written in past tense because you are reporting observations that were made before you started writing the manuscript. The Introduction and Conclusion sections can be a mix of both, but be careful how you marry tenses. If you are writing a statement that is a known fact in the field or some conclusion you are now making from your data, write about these things as if they were always true and in the present tense. (Ex: ADH requires an NAD cofactor for activity.) However, if your wording choice mentions them as a previous finding, the past tense may also be appropriate. (Ex: Cheng and co-authors (#) showed that pyrazole was a competitive inhibitor of ADH.) Sometimes the use of personal pronouns is used in the conclusion section as a way of really owning a claim, but these instances are generally few and far between (Ex: From blah, blah, blah results, we conclude the following model…).

Repetition is necessary. It may seem like you are repeating yourself in various places throughout the manuscript. While absolute monotony is discouraged, repeating key assumptions, rationales or findings in the abstract, introduction, results and conclusions is useful for guiding the reader through your work. Remember, no one else knows the topic as well as you. So, someone reading your work for the first time may not make the necessary connections from rationale to conclusion, especially if those things are only mentioned once and separated by very many sections and experiments. You have to help the reader along or they will get frustrated or start coming to their own alternative conclusions. If this happens during the review process, it means more work for you- re-writing and usually additional experimentation. If it happens after publication, readers will still have doubts about your work.

In this course, you are given small tasks throughout the semester to guide you to the ultimate finished product. Generally, here is the order in which researchers typically work on sections of a manuscript (Note: this is completely different from the order in which they are presented in manuscript form to be submitted to a journal!).

  1. Figures, Tables and Legends: All of that data from your notebooks need to come together in some fashion to tell a story to support your new conclusions. The order of the figures in your manuscript will probably NOT be the order in which you actually did them. Data in figures are used to make the frame for the way you will present your work. I hope your notebook is complete enough to provide the necessary details to render the data into figures. Do you remember what sample was in lane three on that gel you ran 6 months ago? Probably not; hope you wrote it down or you will have to repeat it. If you do have to repeat it, I hope the samples for that gel are in appropriately labeled tubes in a box in the freezer. The figure legends should clearly state what the reader is looking at. Sometimes key details of the method are mentioned when appropriate for evaluating the data. Do not write any lengthy analysis of the results. At this point the researchers that will be authors on the paper will agree upon overall conclusions based on the data and how they will presented in the manuscript.
  2. Title and Author List: The figures should give you an idea of the story you need to tell, so come up with a descriptive, but concise title that captures all of it. Considering the data to be used, the analysis of the results and the writing to be done, authors and their relative ranks in the list according to contribution should be decided.
  3. Materials and Methods: It’s a good section to start with and transition yourself from experimental mode to writing mode. Since you’ve already decided on the figures you need to tell your research story, simply write about the methods you used to generate those figures. Scientific literature varies widely on the useful content within this section. While this section is not a complete protocol, it should give enough highlights of the techniques (and details critical for your experiments) for someone of reasonable ability to repeat your work.
  4. Results: Now that you are in writing-mode, explain all of your results, referring to the data in figures and tables as necessary. This section can be divided into relevant subsections to group findings as they support the various conclusions of your work. It is important to briefly state the purpose for each experiment and the experimental results. Very briefly, conclusions may be mentioned, especially as they lead into later experiments. If this is a pure results section, you won’t want to expound at length on the conclusions or models drawn from your data, but you should say briefly what they mean so it’s not a frustrating cliffhanger for the reader until the next section. Some journals have combined results/conclusions sections and, in that case, it is appropriate to discuss the meanings of your results.
  5. Conclusions: After you have written up all of your results with supporting data in your figures and tables, use this section to talk about what it all means. You may have briefly mentioned the significance of your findings in the results, but here is a chance to put it all together again. This will be the last thing you leave your readers with, so make sure your conclusions are clear and supported by your data. If multiple conclusions are possible, then you’ll have to split the difference and give a little bit of space to each possibility that your experiments haven’t differentiated. Now is the time to relate your conclusions to other published work (which you will cite), supporting or possibly contradictory, as well as offer a possible future direction or application of the research. If you are specifically working on additional experiments along this vein, then it is OK to mention this here or just speak more generally about what your conclusions mean for your field. Research will always keep moving forward; there will always be more experiments to be done, but at some point, you need to stop and write it up. It’s up to you to define the scope of your work. Reviewers may suggest other reasonable experiments, but you can often successfully argue against them if they are beyond the scope you have defined.
  6. Introduction: The background information on your research topic should be sufficient enough for a reasonable person to understand the scope and significance, but not an overly exhaustive review. In theory, it can be written in parallel with your Results and Conclusions sections. It should be complementary to your Results and Conclusions. While it should start with the bigger picture of a research area and its significance, it should ultimately hone in on the problem addressed specifically by your experiments. Usually, the end of the Introduction has a small paragraph describing the current work with respect to techniques and purpose, but not completely giving away the final conclusions. The Introduction and Conclusions sections may need to be edited for content simultaneously so they can most appropriately complement each other.
  7. References: Should be included throughout the writing process- Methods, Results, Conclusions and Introduction. They should include enough information for a read to find the citation and relate it to the claim you have supported in your writing.
  8. Abstract: Take the most important points of the entire work and fit them neatly into a single paragraph of 250 words or less. Your audience should be able to read the abstract and know the gist of what you’re interested in, what you did and the conclusions you made.

Johnna

*Deciding on a journal for submission is a tutorial topic unto itself. I’m avoiding this discussion at this point, since I’ve made the arbitrary decision for my students to prepare according to the Journal of Biological Chemistry.

**If anyone wants to list these rules in the comments section, please do. Gene names are generally italicized, but capitalization/lowercase varies in usage between prokaryotic and eukaryotic systems. As a biochemist, I try to focus on the protein so as not to have to worry about italicization at all.

Suggestions for Science Writing

In the biochemistry lab course I’m teaching, I’m trying to incorporate more scientific communication skills. The basic structure of this lab involves cloning a gene for recombinant protein expression, purifying the protein using affinity chromatography and analyzing the hell out of its kinetics with different substrates and inhibitors. This overall context gives the students insights into the larger scope of a research project instead of discrete, unconnected experiments for each class. It’s challenging enough to make sure the students understand the theory behind all of the techniques and learn some good lab practices along the way. This semester I’m including an extra challenge in terms of scientific writing. Their results will not be confined to their lab notebooks, but will be turned into a manuscript in the style of the Journal of Biological Chemistry.

As part of their lab notebook grade, the students must answer questions related to the experiment of the day. Many of these have been designed to get them to work on their manuscript in a piecemeal fashion throughout the semester (i.e. make this figure, write these methods, etc), so they get feedback from the TAs and myself on all sections before they are assembled into the final manuscript format. It may sound quite organized on paper, but I’ve definitely underestimated the amount of guidance novice authors need to produce quality work. So, I’m trying to come up with a better way of teaching all the things that I’ve learned informally about manuscript-writing over the last decade. I’ll be posting my various tutorials on this blog as a way to further crowdsource useful pro-tips and topics I may not have thought to address. I know many people may have strong opinions on this, so feel free to comment or offer other useful links in the comments section.

Here are the topics/posts/tutorials I’ve got in mind so far…

General Science Writing and Manuscript Preparation

Literature Searching and Reading

Citations and Reference Management

Figures and Figure Legends

Writing Materials and Methods

Writing the Results Section

Writing the Conclusions/Discussion Section

Writing the Introduction

Writing the Abstract

Peer Review and Revisions

My students are mostly seniors on the verge of graduating and moving on to medical and graduate schools or other employment in a health/science field. While they may not all go on to regularly prepare scientific manuscripts, at least they will have a greater appreciation of what it takes. As an exercise, they will get some experience with the process, but I’m not following all of the real life rules. If I did, then the first of my groups to submit would get an A, maybe the second group would get a C and the rest would just fail. And those graded groups would only earn their A or C after an exhausting revision process. For those students choosing to pursue a research career, they’ll learn those harder lessons eventually. As an educational bonus, I’m hoping that working on a manuscript from the side of an author will also allow them to be critical consumers of scientific literature. We should all expect high quality communication of research.

Johnna

References and Links:

https://ugr.ue.ucsc.edu/sites/default/files/jyi_guide_to_scientific_writing.pdf

http://www.studentgroups.ucla.edu/usj/The%20Ins%20and%20Outs%20of%20Scientific%20Manuscript.pdf

http://www.aaeditor.org/StepByStepGuide.pdf

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3626472/pdf/blt-11-217.pdf

On becoming a tree…

“As long as you keep getting born, it’s all right to die sometimes”

Orson Scott Card, The Speaker for the Dead.

I’ve written many times about the differences between autotrophs and heterotrophs on this blog. Loyal readers will know I have a hard-line stance that humans fall decidedly into the heterotroph camp. Nevertheless, some art projects blur the lines between person and plant. So I was more than a little intrigued when a link for the Bios Urn came across my Facebook feed.

BiosUrn

The product offers an alternative to the traditional cemetery as your eternal destination. It’s a special biodegradable urn in which your ashes (or a loved one’s or a pet’s ashes) can be placed along with a tree seed. The whole thing is planted in the ground at a cemetery or other special location. The recycled carbon atoms of your body become the growth medium for a tree that will grow and live on after your death.

biosurn2

It’s beautiful. I get it and as someone who often quotes the Lorax, I’m all for any excuse to plant trees. But honestly, the first thing it made me think of was my all-time favorite book The Speaker for the Dead by Orson Scott Card.* Within the world of this book, humans live on a colony planet that happens to have another species of sentient beings called the Pequeninos. These organisms spend the majority of their life cycle as mammal-like beings, but can pass on into a third life as a tree provided they have been vivisected. It’s pretty gruesome and obviously humans don’t work this way, so cultural misunderstandings ensue.

However, because I’m a stickler for scientific accuracy when it comes to plant science (Hey, someone’s gotta be.), I have one problem with the overly simplistic marketing scheme of the Bios Urn. You don’t actually become the tree. For those of you saying, “I know, I know I won’t be a tree, but in the circle of life my molecules will become part of this tree.” I’ll still be over here at my blog shaking my head, “No, that’s not really how that works.” Here’s a reminder of the photosynthetic equation:

6 CO2 + 6 H2O (+ light) → C6H12O6 + 6 O2

Plants accumulate carbon and mass from atmospheric CO2, not carbon ash or even organic carbon compounds in the soil. The truth is that if you planted your ashes with your tree seed, your carbon would still remain locked in the soil for many years until eventually it is metabolized by microorganisms in the soil and released as CO2 as part of their respiration. This is not a very quick way to turn over your carbon molecules, and your molecules have millennia before they ever become a part of any tree. If you truly wanted your carbon molecules to incorporate into some kind of plant matter, then you need to find a way to carbonate your deceased body. Human carbonated essence could be stored in canisters and then used to supply CO2 to your plant of choice. You could be part of that camellia bush in your front yard, your favorite LSU Oak tree, the General Sherman etc. Scientifically accurate, but good luck marketing that compared to cremation.**

All of this highlights a common misconception about photosynthesis- it’s hard to comprehend how mass can accumulate into living things from air. Our gut tells us mass must come from something more substantial. We memorize the photosynthetic equation in elementary school, but few of us grasp its consequences and really believe it.

“This is how humans are: We question all our beliefs, except for the ones that we really believe in, and those we never think to question.”

Orson Scott Card, The Speaker for the Dead

So, when you die, ask someone to plant a tree or something in your memory. Do it for someone else you’d like to remember, but ashes not really required. Or buy a BiosUrn or put the ashes in a degradable coir starter pot from your local garden center, but you’ll probably want to supplement them with some form of NPK fertilizer. It will help the seed more than your carbon atoms.

Johnna

*It’s a sequel to Ender’s Game for those of you that may not be SciFi freaks.

**Although, in-home soda fountains are a thing now, so if there is a way to carbonate deceased relatives and store them in the handy canisters… well, you get the idea.

Links and References:

https://urnabios.com/

http://en.wikipedia.org/wiki/Speaker_for_the_Dead