Experiment Zero: Using a Raspberry Pi Zero camera

This is the first post at quantixed about Raspberry Pi computing.

Pi Zero is a minimalist Raspberry Pi that can be coupled to a camera. With this little rig, you can make time-lapse footage amongst other things. I’ve set up a couple of these now. One was to make a time-lapse movie of some plants growing through a plastic maze. The results were pretty good and I thought I’d upload the video and a brief how-to guide.

After a delay, you can see four beans sprouting and then one eventually makes it to the top of the maze. This footage was shot over 27 days. The Pi took pictures every 5 min, but I sampled at 10 min in order to make the movie (after discarding the pictures after the sun went down). Everything was automated.

The camera shoots at 3280 × 2464. I downsampled the images to make the video. The camera didn’t focus well on the maze which was a bit too close. Other units are shooting scenery and the autofocus on the unit is great.

How I did it

Pi Zero

Pi Zero with camera module (without IR filter) and a case are available for around £40. I bought mine from the Pi Hut. Power supplies and SD cards are readily available. I put together the PiCam with a fresh Raspbian full image on a 16GB SD card. Another option is to use a smaller card and get the Pi to save the images to a server.

I used PiBaker to format the SD Card, load on Raspbian and add a startup script that would connect the Pi Zero to WiFi and enable VNC. That meant I could plug it in and start using it headless. Well in theory! It turns out that VNC via Mac does not work with the UNIX style password which is the default on the Pi. I needed to connect to a monitor to rectify this by changing to VNC password in the VNC GUI. After this I could log in and use the Pi Zero remotely.

A few more minor steps were needed for full functionality:

  1. I enabled ssh and camera port in Raspberry Pi Configuration, disabled bluetooth and set the correct timezone (this can probably be done in PiBaker but I forgot).
  2. Since I have several Raspberry Pis on the LAN. I needed to give this one its own identity to prevent network conflicts.
  3. I needed to set up SMB sharing on the new Pi.

Instructions for how to do these things are just one google search away.

Now the Pi was ready to start taking images. I built a little stand for it out of Lego and set up the plant maze.

Taking pictures with the Pi

I wrote a shell script to take pictures using raspistill.

I made a directory called camera in home/pi

mkdir camera

Then made a camera.sh file home/pi that looked like this:

#!/bin/bash
DATE=$(date +"%Y-%m-%d_%H%M")
raspistill -o /home/pi/camera/$DATE.jpg

Then I made it executable

chmod +x camera.sh

Using CRON, I execute the shell script on a schedule. I wanted to take pictures every 5 minutes. You can consult cronguru for your needs.

*/5 * * * * /home/pi/camera.sh 2>&1

That’s it! The Pi Zero will happily take pictures until you tell it to stop. Or there’s a…. crash.

Dealing with crashes

If you are going to do long-term time-lapse imaging, you need to defend against a crash that will prevent images from being acquired. In the worst case, the Pi could go offline and you wouldn’t know until you checked up on it. The first one I set up crashed quite often. I couldn’t determine the cause immediately. So I did the next best thing.

I set up a watchdog to monitor for crashes and then reboot the Pi if/when it happens. Many guides online suggest bcm2708_wdog but this doesn’t work for a Pi Zero. Instead this worked for me:

sudo modprobe bcm2835_wdt
sudo nano /etc/modules

Adding the line “bcm2835_wdt” and saving the file

Next I installed the watchdog daemon

sudo apt-get install watchdog chkconfig
chkconfig watchdog on
sudo /etc/init.d/watchdog start
sudo nano /etc/watchdog.conf

I uncommented two lines from this file:

  1. watchdog-device = /dev/watchdog
  2. the line that had max-load-1 in it

Save the watchdog.conf file.

There are guides online that describe how to set up the Pi so that it sends you an email or SMS when there’s a crash/reboot. I figured I didn’t need this – as long as it reboots OK.

What now?

Well, you wait for it to take photos! You can log in via VNC and check that the images are being acquired, or go in via ssh and watch the camera directory fill up. The size of the images is 3280 × 2464 and they are around 4.5 MB each, so the disk can quickly fill.

After a while you’ll want to assemble a movie. I wrote a shell script on my Mac in order to to pull down the images, take a copy of the ones I want and then make a movie file and upload it to Dropbox so I could look at it on the go.

#!/usr/bin/env bash
# move to the location of the images
cd /local/disk/folder2/
# pull down all images to a local folder - only new images are copied
rsync -trv /Volumes/HOMEPI/camera/ /local/disk/folder/
# overnight images are dark and less than 1.5 MB
# copy the ones we want to keep
rsync -trv --min-size=1000K /local/disk/folder/ /local/disk/folder2/
# or you could filter on size like this - delete <2MB
find . -name "*.jpg" -size -2000k -delete
# scale the images down to 480 px wide and make movie
ffmpeg -framerate 30 -pattern_type glob -i '*.jpg' -c:v libx264 -pix_fmt yuv420p -vf scale=480:-2 out.mp4
# move to dropbox
mv out.mp4 /My/Dropbox/Folder/out.mp4

This script means that I had to manually delete the pictures from the Pi once they’d been copied but that was OK. My plan is to write a script to do this for the longer running projects so that it is automated.

While it is possible to make the movies on the Pi itself, I did it on the Mac as that computer is beefier and is not busy taking pictures every 5 min! ffmpeg is a great tool for this and the documentation is impressive. For example if you have set up the camera in the wrong orientation you can do transposition in ffmpeg. If you don’t have ffmpeg, it is a simple install on the command line.

ruby -e "$(curl -fsSL https://raw.githubusercontent.com/Homebrew/install/master/install)" < /dev/null 2> /dev/null
brew install ffmpeg

Hopefully this guide is useful to you. The Pi Zero Camera can be used for streaming video as well as taking a series of still images. I’m planning to test this out soon.

—

The post title “Experiment Zero” comes from the title of the album by Man or Astro-Man?

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Til I Die: Seeking new music

I’ve been following the tweets from an account called Albums You Must Hear @Albums2Hear. Each tweet is an album recommended by the account owner. I’m a sucker for lists of Albums That I Must Hear Before I Die since I’m always interested in new (or not so new) music recommendations.

I wanted to assemble a list of the albums that I don’t have from this account and I was able to do so using R.

Using rtweet, it was possible to pull a list of all the albums and reorganise them so that I had a csv containing the albums with the artist and year. I could then use this to compare with a list of albums from my iTunes library. A snippet of the retrieved records is shown here (full list is here).

The code for retrieval is here. The output is csv can be used to compare with a list of your own records.


library(rtweet)
library(httpuv)
library(stringr)
all_tweets <- get_timeline("Albums2Hear", n = 1500)
albums <- all_tweets$text
albums <- gsub("#albumsyoumusthear ","",albums)
tempdf <- as.data.frame(str_split_fixed(albumV, " - ", 3))
colnames(tempdf) <- c("Artist","Album","YearURL")
tempdf2 <- as.data.frame(str_split_fixed(tempdf$YearURL, " ", 2))
colnames(tempdf2) <- c("Year","URL")
df <- data.frame(y$Artist,y$Album,z$Year)
colnames(df) <- c("Artist","Album","Year")
write.csv(df,file = "albums2hear.csv")

Thanks to whoever runs the account – they ask for support here.

—

The post title comes from ‘Til I Die by The Beach Boys from Sunflower/Surf’s Up.

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Tips from the blog XI: docx to pdf

A long time ago I posted a little Automator routine to convert Word doc/docx files to PDF. Not long after that, this routine ceased to work due to changes in Microsoft Word (I think). It’s still very useful to convert a whole folder of docx files to PDF in order to avoid Word and just use Preview on the Mac. For committee work or for marking students’ work, I often have a whole folder of docx files and would prefer it if they were in PDF format. I found this very nifty trick on the web and thought I’d post a link here to make up for the fact that my old post no longer works.

The full post is here. What is so nice about this Automator solution is that it uses a bash script to do the conversion. This means you don’t need Microsoft Word for it to work! From what I can see it uses the xml in the docx file (and presumably won’t work on older *.doc files) for the conversion. The post describes how to run it as a Service in macOS. Note, that it destroys the docx files, so it should only be used on a copy. It could be run from the command line rather than right-clink, the engine is this little script.

Thanks to Jacob Salmela for posting it.

—

This post is part of a series of short tips

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New Lexicon: how to add a custom minted lexer in Overleaf

This quick post comes courtesy of LianTze Lim (an Overleaf TeXpert) and Kota Miura (a bioimage analyst).

I asked on the ImageJ forum some time ago how to add an ImageJ Macro lexer for a LaTeX document I was writing. Kota responded with this lexer for pygments. I then asked Overleaf if it was possible to add a custom lexer to an Overleaf document using the minted package. At the time this was not possible. However, I got a message from them today with a solution.

Steps to do this for your own Overleaf project:

  1. Add Kota’s imagejmacro.py file to your project
  2. Add minted to your preamble and then use
\begin{minted}{imagejmacro.py:ImageJMacroLexer -x}
// your code
\end{minted}

Here, imagejmacro.py is the name of the custom lexer saved in your project and ImageJMacroLexer is the name of the class in that file. If you want to use another custom lexer just replace as required. I have put up a read-only Overleaf example to show it working.

Thanks to LianTze for following up with me about this and special thanks to Kota who wrote the custom lexer.

—

The post title comes from the LP of the same name by Paint It Black.

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Pledging My Time IV

 

The Green Leek 10.5 km run is a mixed terrain race now in its third year. Today’s was a wet and muddy edition. The chip times were posted this afternoon and using my previous code, I took a look at the results.

I was a bit disappointed with my time, which was about 24 s slower than last year. Considering that I’m running faster this year than last, I wondered whether the conditions affected my time. To look at this I quickly retrieved times for people who’d run it all three editions and looked to see if this edition was generally slower than previous editions.

Excuse the formatting of the plot. It looked pretty flat but then we’re probably only considering very small differences over 10.5 km. So I looked at the difference in time from the 2016 edition. Again the formatting is bad (23:55 is 5 minutes faster than 2016, 00:05 is 5 minutes slower).

Three people recorded much slower times this year, but the majority are within the difference from 2016 to 2017. Obviously this is just a few people that could be easily picked out using a script, more runners might reveal more of a pattern. Anyway, here’s hoping for better weather next year!

Well done to Andy Crabtree and Rachel Miller who were fastest male and female, respectively. Thanks to the organisers and volunteers.

—

The post title is taken from “Pledging My Time” a track from Blonde on Blonde by Bob Dylan

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Multiplex: Small multiple artwork from GPX tracks

I’d seen the small multiple artwork of running and cycling routes from Marcus Volz’s R package Strava all over the web. Ads for “posters of your GPS tracks” pop up on Reddit and I’d notice a few #Rstats people put up their posters on Twitter. I’ve had the package bookmarked for a while and this week I finally got round to generating a small multiple poster of some of my cycling routes.

I was pleased with the result and wanted to post it here. But also, running the code was not straightforward as I’ll explain below. If you want to generate your own plot read on.

The idea behind the poster is really nice. You get a kind of generative art-style poster. It looks nice and you can identify individual routes which is fun to do.

The instructions on the GitHub page are absolutely correct and the code should run out-of-the-box. The idea is that you download your Strava data and then make your plots. Unfortunately, it seems that a change in Strava’s data export policy (possibly related to GDPR changes) has broken the package. I found that there are two problems. First, Strava’s “download your data” link gives you a mix of formats (in my case GPX and FIT files), the package only works with GPX. Second, if there is any elevation data missing from a track, the data frame that is needed to make the poster is not built properly.

Going GPX only: In my case, I don’t keep all my data in Strava and instead use a local repository managed with RubiTrack. This software allows me to filter for the tracks I want and export them in GPX format. The only problem is that it generates one huge file with all the tracks enclosed. This gets read by the package as a single track. To fix this, I split the file using awk.

awk '/<trk>/{close(file);n++;}{file="track"n".gpx";print >> file;}' untitled.gpx

I could then discard track1.gpx which just had the xml header and then use the directory of gpx files.

The elevation problem: this affected only some of the tracks, so in the end the R code needed to be modified. The elevation data is not needed to make the posters so the file process_data.R needs editing, line 28 can be commented out and then line 32 should read:

result <- data.frame(lat = lat, lon = lon, time = time, type = type) %<%

This issue is raised on GitHub and has been closed, but the code doesn’t work with elevation blanks. If you run into this problem, this is the way I found to fix it. The other plots in the package which do use elevation will not run, but at least the poster can be made.

I exported the poster as PDF and then made some changes in Illustrator to give the result above.

—

The post title comes from Multiplex from Oliver’s Standing Stone LP from 1974.

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My Blank Pages VI: Programming in Igor Pro

It has been a long time since I wrote a book review.

A few months ago I read on IgorExchange that Martin Schmid had written a book about programming Igor. I snapped up a copy. I’m a competent Igor programmer but I was hoping that this book would be useful for lab members that want to learn.

Learning Igor – like most IDEs or programming languages – is tough going. There’s a booklet from WaveMetrics (the company that sells Igor Pro) called Getting Started – which is really good. There are a few other guides on the web (Payam’s guide, Thomas Braun’s coding conventions, quantixed’s own translator), but other resources are pretty scarce. The Igor Manual itself is excellent but it’s many, many pages long and is only meant to be consulted. So I was intrigued whether Martin Schmid’s book would fill the gap between Getting Started and more advanced guides.

What makes Igor Pro so fantastic is the way that you can use it for so many different things: image processing, statistics, graphing, curve fitting, instrument control and so on. Part of the challenge of writing a book on Igor Programming is deciding what to cover. Schmid deals with this by covering basic programming and core-intermediate topics such as dialogs, loops, string magic etc. The book stops short of any specialised applications. So it’s a really useful intermediate programming guide. It’s a great little book and is recommended for those who want to dig further after doing the Getting Started exercises.

I knew I would learn something from the book because there’s always alternative ways to do stuff in Igor: things that you didn’t know about or little tricks to do stuff faster. What I was surprised about was the first thing mentioned in the book was new to me. The author favours module-static programming. All of my Igor programming has been done in the global pragma and I have avoided this more C-like way of encapsulating programs that I’ve written so far. Module-static works well because it eliminates naming conflicts. I have dealt with name conflicts by using static functions which are called from the top of the stack, and the top has a unique name (arguably this is the same as module-static, but not identical). As the Igor Manual says “this gets tedious after a while” and that’s true. Although in my defence, name conflicts are generally not a problem for the way I work because I favour a reproducible approach. A new experiment is started – one user-written ipf is opened – and the code is run. This means naming conflicts are minimised. The book has actually convinced me that module-static is a good thing, especially since my Igor code is now deployed around the lab and naming conflicts could easily become a problem. It’s an advanced programming technique but is dealt with early by the author and it kind of works. After this, more basic programming topics are covered in depth.

There’s always room for improvement: there are several example programs at the back which need to be rekeyed to run, since this is a paper book and no electronic version is available AFAIK. The author has put one up here to save rekeying and another here, but otherwise you need to type in the examples to see what will happen. This is too long-winded. I’ve been spoiled reading texts about R where the examples can all be run from a markdown file inside RStudio. It would’ve been nice if the code was made available for this book. I don’t think it would compromise the value of the book since it is the text that is most valuable.

The book is available at Amazon for £7.99 at the time of writing.

—

My Blank Pages is a track by Velvet Crush. This is an occasional series of book reviews.

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Pledging My Time III

I’ve previously crunched times for local Half and Full Marathons here on quantixed. Last weekend was the Kenilworth Half Marathon (2018) over a new course. I thought I’d have a look at the distributions of times and paces of the runners. The times are available here. If the Time and Category for finishers are saved as a csv, the script below works to generate the following plots.

Aggregated stats for the race are here. The beeswarm plot nicely shows the distribution of runners times and paces per category. There’s a bimodality to some of the age groups which is interesting. You can see from the average times that people get slower as they get older, as expected.

There was a roughly 2:1 split of M:F runners with a similar proportion in all categories. The ratio is similar for DNSers. The winning times were Andrew Savery of Leamington C A & C in MV35 with 01:12:51 and Polly Keen of Nuneaton Harriers in F sen with 01:23:46.

Congrats to everyone who ran and thanks to the organisers and all the supporters out on the course.


require(ggplot2)
require(ggbeeswarm)
file_name <- file.choose()
df1 <- read.csv(file_name, header = TRUE, stringsAsFactors = FALSE)
# aggregate M and F to a new category called Gender
df1$Gender <- ifelse(startsWith(df1$Category,"F"),"F","M")
# format Date column to POSIXct
df1$Time <- as.POSIXct(strptime(df1$Time, format = "%H:%M:%S"))
orig_var <- as.POSIXct("00:00:00", format = "%H:%M:%S")
p1 <- ggplot( data = df1, aes(x = Category,y = Time, color = Category)) + 
  geom_quasirandom(alpha = 0.5, stroke = 0) +
  stat_summary(fun.y = mean, geom = "point", size=2, aes(group = 1)) +
  scale_y_datetime(date_labels = "%H:%M:%S", limits = c(orig_var,NA))
p1
# instead of finishing time, let's look at pace (min/km)
df1$Pace <- as.numeric(difftime(df1$Time, orig_var) / 21.1) * 3600
df1$Pace <- as.POSIXct(df1$Pace, origin = orig_var, format = "%H:%M:%S")
p2 <- ggplot( data = df1, aes(x = Category,y = Pace, color = Category)) + 
  geom_quasirandom(alpha = 0.5, stroke = 0) +
  stat_summary(fun.y = mean, geom = "point", size=2, aes(group = 1)) +
  scale_y_datetime(date_labels = "%M:%S", limits = c(orig_var,NA))
p2
# calculate speeds rather than pace
df1$Speed <- 21.1 / as.numeric(difftime(df1$Time, orig_var))
p3 <- ggplot( data = df1, aes(x = Category, y = Speed, color = Category)) + 
  geom_quasirandom(alpha = 0.5, stroke = 0) +
  stat_summary(fun.y = mean, geom = "point", size=2, aes(group = 1)) +
  ylim(0,NA) + ylab("Speed (km/h)")
p3
# now make the same plots but by Gender rather than Category
p4 <- ggplot( data = df1, aes(x = Gender,y = Time, color = Gender)) + 
  geom_quasirandom(alpha = 0.5, stroke = 0) +
  stat_summary(fun.y = mean, geom = "point", size=2, aes(group = 1)) +
  scale_y_datetime(date_labels = "%H:%M:%S", limits = c(orig_var,NA))
p4
p5 <- ggplot( data = df1, aes(x = Gender,y = Pace, color = Gender)) + 
  geom_quasirandom(alpha = 0.5, stroke = 0) +
  stat_summary(fun.y = mean, geom = "point", size=2, aes(group = 1)) +
  scale_y_datetime(date_labels = "%M:%S", limits = c(orig_var,NA))
p5
p6 <- ggplot( data = df1, aes(x = Gender, y = Speed, color = Gender)) + 
  geom_quasirandom(alpha = 0.5, stroke = 0) +
  stat_summary(fun.y = mean, geom = "point", size=2, aes(group = 1)) +
  ylim(0,NA) + ylab("Speed (km/h)")
p6
ggsave("raceTimeByCat.png", plot = p1)
ggsave("racePaceByCat.png", plot = p2)
ggsave("raceSpeedByCat.png", plot = p3)
ggsave("raceTimeByGen.png", plot = p4)
ggsave("racePaceByGen.png", plot = p5)
ggsave("raceSpeedByGen.png", plot = p6)

Edit 2018-09-12T18:52:43Z I wasn’t happy with the plots and added a few more lines to look at gender as well as category. And show speed as well as pace and finishing time.

—

The post title is taken from “Pledging My Time” a track from Blonde on Blonde by Bob Dylan

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Rip It Up: Grabbing movies from Twitter for use in ImageJ

Some great scientific data gets posted on Twitter. Sometimes I want to take a closer look and this post describes a strategy to do so.

Edit: I received a request to take down the 3D volume images derived from the example dataset I used in this post. I’ve edited the post below so that is now a general guide.

Grab the video

It can be a bit difficult to the grab video from Twitter. The best way I’ve found is using youtube-dl. This works for downloading video and audio from YouTube to view offline, but it also works for other embedded video content on other websites.

To download the video use:

youtube-dl -o '%(title)s.%(ext)s' https://twitter.com/username/status/tweetID

this downloads an mp4 file which is automatically named.

Convert to avi

Now, mp4 is a compressed file format which cannot be read directly by FIJI/ImageJ. Conversion to avi means that the file can be loaded. I like to use another command line tool, ffmpeg for video conversions.

ffmpeg -i originalFile.mp4 -pix_fmt nv12 -f avi -vcodec rawvideo convertedFile.avi

Now we have an avi file called convertedFile.avi that we can use.

Load into FIJI

The avi can be loaded into FIJI. At this point you can analyse the video. However, in the case of the video I was interested in, the data had been pseudocolored and was now in RGB format. I wanted to look at the original data. Converting to grayscale does not give the correct representation but conversion back to grayscale is possible if you know the LUT was applied. Even if you don’t, it’s possible to take a guess at the LUT and do the conversion.

Converting RGB to original values

I found a nice gist that does the conversion for a single image. I just modified this code to work for a stack. It requires the LUT to be displayed vertically in a window called LUT. Caution: this code runs very slowly because every pixel in every slice needs to be recalculated and ImageJ is slow… I took a guess that mpl-inferno was used (I don’t think is exactly right but it worked well enough). You can display the built-in LUTs in FIJI using Color > Display LUTs… and from there you can make the LUT window which the macro uses for the calculation. The macro to convert stacks to grayscale using the LUT is here.

I had a nice grayscale version of the data (inverted because I wanted to look at the volume). This let me see how the layers in the original video add together to make the full structure. I used ClearVolume which can be installed via Update Site in FIJI. I just made a quick video to show it in action (see below). You’ll have to take my word for it (video removed).

So extracting scientific data from Twitter or another online source is pretty straightforward. The extra complication was getting rid of the pseudocoloring, but once this was done, something very close to the original data was available.  Nonetheless this workflow is a fun way to take a closer look at some of the cool movies that people post on Twitter. I hope you find it useful.

—

The post title comes from “Rip It Up” by Orange Juice. A popular title in my library with versions from several different artists. I was thinking what is described is similar to ripping video content.

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Ferrous: new paper on FerriTagging proteins in cells

We have a new paper out. It’s not exactly news, because the paper has been up on bioRxiv since December 2016 and hasn’t changed too much. All of the work was done by Nick Clarke when he was a PhD student in the lab. This post is to explain our new paper to a general audience.

The paper in a nutshell

We have invented a new way to tag proteins in living cells so that you can see them by light microscopy and by electron microscopy.

Why would you want to do that?

Proteins do almost all of the jobs in cells that scientists want to study. We can learn a lot about how proteins work by simply watching them down the microscope. We want to know their precise location. Light microscopy means that the cells are alive and we can watch the proteins move around. It’s a great method but it has low resolution, so seeing a protein’s precise location is not possible. We can overcome this limitation by using electron microscopy. This gives us higher resolution, but the proteins are stuck in one location. When we correlate images from one microscope to the other, we can watch proteins move and then look at them with high resolution. All we need is a way to see the proteins so that they can be seen in both types of microscope. We do this with tagging.

Tagging proteins so that we can see them by light microscopy is easy. A widely used method is to use a fluorescent protein such as GFP. We can’t see GFP in the electron microscope (EM) so we need another method. Again, there are several tags available but they all have drawbacks. They are not precise enough, or they don’t work on single proteins. So we came up with a new one and fused it with a fluorescent protein.

What is your EM tag?

We call it FerriTag. It is based on Ferritin which is a large protein shell that cells use to store iron. Because iron scatters electrons, this protein shell can be seen by EM as a particle. There was a problem though. If Ferritin is fused to a protein, we end up with a mush. So, we changed Ferritin so that it could be attached to the protein of interest by using a drug. This meant that we could put the FerriTag onto the protein we want to image in a few seconds. In the picture on the right you can see how this works to FerriTag clathrin, a component of vesicles in cells.

We can watch the tagging process happening in cells before looking by EM. The movie on the right shows green spots (clathrin-coated pits in a living cell) turning orange/yellow when we do FerriTagging. The cool thing about FerriTag is that it is genetically encoded. That means that we get the cell to make the tag itself and we don’t have to put it in from outside which would damage the cell.

What can you use FerriTag for?

Well, it can be used to tag many proteins in cells. We wanted to precisely localise a protein called HIP1R which links clathrin-coated pits to the cytoskeleton. We FerriTagged HIP1R and carried out what we call “contextual nanoscale mapping”. This is just a fancy way of saying that we could find the FerriTagged HIP1R and map where it is relative to the clathrin-coated pit. This allowed us to see that HIP1R is found at the pit and surrounding membrane. We could even see small changes in the shape of HIP1R in the different locations.

We’re using FerriTag for lots of projects. Our motivation to make FerriTag was so that we could look at proteins that are important for cell division and this is what we are doing now.

Is the work freely available?

Yes! The paper is available here under CC-BY licence. All of the code we wrote to analyse the data and run computer simulations is available here. All of the plasmids needed to do FerriTagging are available from Addgene (a non-profit company, there is a small fee) so that anyone can use them in the lab to FerriTag their favourite protein.

How long did it take to do this project?

Nick worked for four years on this project. Our first attempt at using ribosomes to tag proteins failed, but Nick then managed to get Ferritin working as a tag. This paper has broken our lab record for longest publication delay from first submission to final publication. The diagram below tells the whole saga.

 

The publication process was frustratingly slow. It took a few months to write the paper and then we submitted to the first journal after Christmas 2016. We got a rapid desk rejection and sent the paper to another journal and it went out for review. We had two positive referees and one negative one, but we felt we could address the comments and checked with the journal who said that they would consider a revised paper as an appeal. We did some work and resubmitted the paper. Almost six months after first submission the paper was rejected, but with the offer of a rapid (ha!) publication at Nature Communications using the peer review file from the other journal.

Hindsight is a wonderful thing but I now regret agreeing to transfer the paper to Nature Communications. It was far from rapid. They drafted in a new reviewer who came with a list of new questions, as well as being slow to respond. Sure, a huge chunk of the delay was caused by us doing revision experiments (the revisions took longer than they should because Nick defended his PhD, was working on other projects and also became a parent). However, the journal was really slow. The Editor assigned to our paper left the journal which didn’t help and the reviewer they drafted in was slow to respond each time (6 and 7 weeks, respectively). Particularly at the end, after the paper was ‘accepted in principle’ it took them three weeks to actually accept the paper (seemingly a week to figure out what a bib file is and another to ask us something about chi-squared tests). Then a further three weeks to send us the proofs, and then another three weeks until publication. You can see from the graphic that we sent back the paper in the third week of February and only incurred a 9-day delay ourselves, yet the paper was not published until July.

Did the paper improve as a result of this process? Yes and no. We actually added some things in the first revision cycle (for Journal #2) that got removed in subsequent peer review cycles! And the message in the final paper is exactly the same as the version on bioRxiv, posted 18 months previously. So in that sense, no it didn’t. It wasn’t all a total waste of time though, the extra reviewer convinced us to add some new analysis which made the paper more convincing in the end. Was this worth an 18-month delay? You can download our paper and the preprint and judge for yourself.

Were we unlucky with this slow experience? Maybe, but I know other authors who’ve had similar (and worse) experiences at this journal. As described in a previous post, the publication lag times are getting longer at Nature Communications. This suggests that our lengthy wait is not unique.

There’s lots to like about this journal:

  • It is open access.
  • It has the Nature branding (which, like it or not, impresses many people).
  • Peer review file is available
  • The papers look great (in print and online).

But there are downsides too.

  • The APC for each paper is £3300 ($5200). Obviously open access must cost something, but there a cheaper OA journals available (albeit without the Nature branding).
  • Ironically, paying a premium for this reputation is complicated since the journal covers a wide range of science and its kudos varies depending on subfield.
  • It’s also slow, and especially so when you consider that papers have often transferred here from somewhere else.
  • It’s essentially a mega journal, so your paper doesn’t get the same exposure as it would in a community-focused journal.
  • There’s the whole ReadCube/SpringerNature thing…

Overall it was a negative publication experience with this paper. Transferring a paper along with the peer review file to another journal has worked out well for us recently and has been rapid, but not this time. Please leave a comment particularly if you’ve had a positive experience and redress the balance.

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The post title comes from “Ferrous” by Circle from their album Meronia.

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