eamag/protocols-with-steps huggingface dataset:
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
This protocol describes a wet-lab method to identify Leishmania proteins bands recognized by human IgG1. The cytosolic proteins obtained by cell lysis are initially separated by tricine gel electrophoresis using a long (14 cm) apparatus (SG-200, CBS Scientific, USA). Upon completion of the gel electrophoresis, the proteins are transferred onto a nitrocellulose (NC) membrane (10600046, GE Healthcare Life Sciences, UK) using a semi-dry electroblotting transfer system (1703940, Bio-Rad, UK). The NC membrane is sliced into several thin (4mm) strips and each of the strip is immunoassayed with individual sera from Indian patients with different clinical status of visceral leishmaniasis - VL (active VL, relapse, endemic healthy controls and non-endemic healthy controls) using an HRP-conjugated anti-human IgG1 secondary antibody (ab99774, Abcam, Cambridge, UK)

[STEPS]
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein transfer
?. 
SECTION: Protein transfer
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein separation by tricine SDS-PAGE
?. 
SECTION: Protein transfer
?. 
SECTION: Protein transfer
?. 
SECTION: Protein transfer
?. 
SECTION: Protein transfer
?. 
SECTION: Protein transfer
?. 
SECTION: Protein transfer
?. 
SECTION: Protein transfer
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?. 
SECTION: Immunoassay
?.
TITLE: Rapid Protocol for Single Sporing Ascochyta rabiei
AUTHORS: Melody Christie, Kevin Moore

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>A rapid protocol for single sporing of </span><span style = "font-style:italic;">A. rabiei.</span><span> This protocol reduces the time required for single sporing to just 48 hours and allows rapid detection of contamination.</span><span style = "font-style:italic;"> </span></div></div>

[STEPS]
?. Pour water agar plates (~ 2 cm thickness and containing antibiotic) 1-2 days before culturing to ensure plates are dry.
?. Autoclave 20 ml glass bottles (McCartney) containing 10 ml milliQ water with aluminum lids loosely on. Allow to cool to room temperature.
?. Add ~ 2 drops of  (approximately ) to each bottle.
30 µl
?. Place a short segment of stem with lesion and visible pycnidia into bottle, tighten lid and shake to mix.
?. Incubate for  at room temperature ().
25 °C
?. Vigorously shake McCartney bottle to release spores into suspension (shake for ).
?. Pour suspension on to prepared media and immediately pour off.
?. Seal petri dishes with parafilm and incubate at room temperature ().
25 °C
?. Examine conidia development/spore germination between 24 -  incubation for single spore isolation.
?. Adjust the focus point of the dissecting microscope to the surface of the media to find germinating spores.
?. A small piece of media with the target spore attached should be picked up using a sterilized needle and transferred to 90 mm V8 media plate (containing antibiotic).  Place 3-4 individual spores on each plate evenly.
?. When placing spore on media produce a small cut as you set it down.  This will concentrate any bacterial contamination inside the cut and make it easy to avoid during spore isolation (see attached image).
TITLE: Breath Analysis System for Detecting Breath Pattern Signature of COVID-19 
AUTHORS: Salvatore D Morgera, Tiffany Miller,  Arash Takshi, Stephen E Saddow, Matthew Palm

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">The magnitude of how rapidly the COVID-19 virus could spread and infect others facilitated the need for a rapid COVID-19 test, accessible and affordable for both children and adults worldwide. As a result, an infected patient can quickly quarantine and isolate to slow the spread. Our research group has determined VOCs associated with COVID-19 and has built a prototype gas analysis system similar to a breathalyzer for detecting gasses of COVID-19. Some of the symptoms like headaches, coughing, and diarrhea result in an inflammatory response to create a unique variety of gasses like acetone, hydrogen, and carbon monoxide that can be detected when they are exhaled through a person’s breath. A sample of a person’s breath is first collected in a chamber when a patient breaths into the valve, the gas sensors then detect a voltage change when they contact a COVID-19 target gas. The concentration in parts-per-million is calculated based on the non-linear resistance ratio of the target gas and clean air. Log-based scale calculations and post-data processing calibrate the sensors to increase the accuracy and selectivity. Finally, the voltages and PPM of the target gas concentration data is stored in a computer database and sensor coding reveals the resulting breath pattern signature of COVID-19. At just under $15.00 United States Dollars (USD), test results are just a breath away, within seconds of taking the test and even before leaving the doctor’s office. Although, this system may also be modified to sniff out other pathogens of disease, spoiled meats, and ripened fruit, we are dedicated to build a gas sensor platform for low cost, noninvasive, and rapid detection of the breath pattern signature of COVID-19, in an attempt to slow the spread of the global health threat of infection and in turn to help save lives.</div><div class = "text-block">Breath analysis systems serve as a noninvasive means for disease screening. Our objective is to utilize this prototype to obtain a database of H2, CO, NO, Acetone and Alcohol concentrations, whereby, COVID patient breath samples are compared to healthy individual breath samples using our breath analysis system prototype. This system is configured for measuring the aforementioned gas concentrations in ppm and recording the data to a text file in a computer database for subsequent analysis. The data obtained from this study is critical to the development of this system.</div></div>

[STEPS]
TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>Atmospheric methane is rapidly lost when it enters humid subterranean critical and vadose zones (e.g., air in soils and caves). Because methane is a source of carbon and energy, it can be consumed by methanotrophic methane-oxidizing bacteria. As an additional subterranean sink, it has been hypothesized that methane is oxidized by natural radioactivity-induced radiolysis that produces energetic ions and radicals, which then trigger abiotic oxidation and consumption of methane within a few hours. Using controlled laboratory experiments, we tested whether radiolysis could rapidly oxidize methane in sealed air with different relative humidities while being exposed to elevated levels of radiation (more than 535 kBq m<sup>-3</sup>) from radon isotopes <sup>222</sup>Rn and <sup>220</sup>Rn (i.e., thoron). We found no evidence that radiolysis contributed to methane oxidation. In contrast, experiments with moist soils in the same apparatus without added radon isotopes confirmed biodegradation of methane by methane-oxidizing bacteria.</p>

[STEPS]
SECTION: Apparatus at Indiana University for active, time-resolved measurements of gas concentrations with circular flow
?. 
SECTION: Discriminating between α-radiation from radon Rn-222 versus thoron Rn-220 and their progeny at Indiana University
?. 
SECTION: Basics of active time-series measurements at Indiana University
?. 
SECTION: Humidity and temperature effects at Indiana University
?. 
SECTION: Influence of radiation and Methane-Oxidizing-Bacteria (MOB) on methane loss at Indiana University
?. 
SECTION: Gas-tight terrarium experiments at Royal Holloway University of London
?. 
SECTION: Overview of experimental approaches
?.
TITLE: plasma preparation_test
AUTHORS: Anneleen Decock

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol describes how to prepare plasma from blood and is an experimental protocol.</div></div>

[STEPS]
?. [Blood draw]
Note blood tube typeNote date and time point of blood collection; store tubes upright at room temperature until centrifugationNote date and time point of arrival in labNote amount of blood that was collectedTake pictures of the tubes if needed
?. [Centrifugation step 1: 20 min at 400 g (rcf)]
Invert tubes 5 times before starting centrifugationSpin tubes for 20 min at 400 g (rcf) (without brake), at room temperaturePipette platelet-rich plasma (PRP) carefully into a new collection tube, leave ± 0.5 cm above the buffy coat (do not disturb the buffy coat)Aliquot the PRP into cryovials or LoBind tubes, snap freeze in liquid nitrogen and store at -80°C (note time point in freezer), or continue to prepare platelet-poor plasma (PPP)
?. [Centrifugation step 2: 10 min at 800 g (rcf)]
Spin the PRP for 10 min at 800 g (rcf) (without brake) to obtain platelet-poor plasma (PPP), at room temperaturePipette PPP carefully into a new collection tube, leave ± 0.5 cm above pellet (do not disturb pellet)Optional: If platelets need to be collected, resuspend pellet in remaining volume above pellet, transfer to 1.5 ml tube, and spin again for 10 min at 800 g (rcf) (without brake), remove volume and snap freeze pellet in 1.5 ml tube. 
Aliquot the PPP into cryovials or LoBind tubes, snap freeze in liquid nitrogen and store at -80°C (note time point in freezer), or continue to prepare platelet-free plasma (PFP)
?. [Centrifugation step 3: 15 min at 2500 g (rcf)]
Spin the PPP for 15 min at 2500 g (rcf) (without brake) to obtain platelet-free plasma (PFP), at room temperaturePipette PFP carefully into a new collection tube, leave ± 0.5 cm above pellet (do not disturb pellet)Aliquot the PFP into cryovials or LoBind tubes, snap freeze in liquid nitrogen and store at -80°C (note time point in freezer)
TITLE: Coffee Protocol
AUTHORS: John   Borghi

[DESCRIPTION]
This protocol is intended as a demonstration of what I like to call the Coffee Cup Rule - Document more than you think is necessary and with greater specificity that you think is necessary.

The name is derived from the fact that the cup measures on drip coffee machines refer to "cups of coffee" a unit of measure that is between 4 and 6 fluid ounces rather than a "cup", which is 8 fluid ounces in the United States. Because of this, it is important to be very specific when describing the amounts needed to make a pot of coffee. 

The directions here will make a good enough cup of coffee, but a truly replicable protocol would require even more specificity about the roast of the coffee, the grind of the coffee, the temperature of the water, the make and model of the drip coffee machine, etc. The amounts and directions would also be different if using other methods for brewing coffee. If you're using a French Press, you'd probably want a slightly different coffee-to-water ratio than what's described here.

[STEPS]
SECTION: Make Coffee
1. Measure out 1000 g and 60 g. 

If the scale is out of batteries, 4 1/4 cups (US) of water and 8 tablespoons of coffee will work in a pinch.  

Use a measuring cup for the water not the cup measures on the side of the coffee pot. Those refer to "cups of coffee".
SECTION: Make Coffee
2. Pour the water in the drip coffee machine. 

Pour the coffee into the filter and put the filter into the machine. 

Brew on "Bold" setting, if available.
SECTION: Make Cup of Coffee
3. Pour  30 g and 200 g in a mug.

Enjoy!
TITLE: Sinai SCENT TMC - Peripheral Blood Collection
AUTHORS: Santos Bermejo, Sojin Kim, Patty J Lee

[DESCRIPTION]
This Standard Operating Procedure is to provide guidance toresearch team members who are involved in the retrieval, processing, and storing of peripheral blood samples obtained from research participants.

The research team shall use this SOP within the scope of an IRB-approved protocol.

[STEPS]
SECTION: Scope
1. This document outlines the process of collecting, processing, and storing peripheral blood from research participants enrolled in SCENT U54. 
SECTION: Materials
2. Blood Collection 
2x Yellow Top Tubes (PBMC) - Acid citric dextrose additive (ACD) 
1x Red Top Tube (Serum) – No additive 
3x Purple Top (Plasma) - Ethylenediaminetetraacetic Acid (EDTA) 
1x PaxGene Tube 
Blood Collection Supplies: Gloves, Butterfly needle, vacutainer, tourniquet, gauze, alcohol pads, Band-Aids 
Biohazard bag 
Transport container (check with lab manager for appropriate transport container)
SECTION: Procedures - Blood collection and Transport
4. Sample retrieval:  
Peripheral blood shall be collected during visit 2, immediately before the bronchoscopy procedure.
SECTION: Materials
3. Processing 
Microtubes (5x 0.2mL, 1-2x 1.5mL) 
PBS (Phosphate Buffered Saline)
Histopaque – 1077 (Sigma, 10771) 
10% bleach solution (to discard used tips, tubes, etc.)
SECTION: Procedures - Blood collection and Transport
5. Order of blood draw:  
Red → Yellow → Purple → PaxGene
SECTION: Procedures - Blood collection and Transport
6. Inversion times
Once blood has stopped flowing in the tube, remove and gently invert (Avoid vigorous shaking or mixing): 
Red – No inversion necessary 
Yellow – 8 times 
Purple – 8 times 
PaxGene – 8 times. Do not force blood into the tube, which can affect RNA quality.
SECTION: Procedures - Blood collection and Transport
7. Sample Transport: 
Samples should be transported to the lab and processed 30-60 minutes after collection. Transport should occur at Room temperature 

SECTION: Procedures - Sample Processing
8. Serum (Red top) and Plasma (Purple top):  

Label all tubes with Study ID and ‘S’ for serum, and ‘P’ for plasma. 
Centrifuge tubes 2200 rpm, 10 min 
After spinning, aliquot supernatant to 5 x 0.2mL microtubes, and the remaining supernatant into 1.5mL tubes (label top of 1.5mL w/ volume (e.g. ‘300cc’) 
Store in -80 °C freezer
Update sample inventory

SECTION: Procedures - Sample Processing
9. PaxGene:  

Store the PAXgene Blood RNA Tube upright Room temperature (18°C–25°C) for a minimum of 2 hours and a maximum of 72 hours before processing or transferring to the refrigerator (2–8°C) or freezer (−20°C). 
Update sample inventory
SECTION: Procedures - Sample Processing
10. PBMC Isolation and Cryopreservation: 

See PBMC isolation protocol in LRT (Lee Research Team) shared drive
TITLE: AFLP RedTaq protocol
AUTHORS: Michal Ronikier, Tomasz Suchan

[DESCRIPTION]
<div class = "text-blocks"></div>

[STEPS]
?. [Restriction-ligation reaction]
[water]
[T4 DNA Ligase buffer (10×)]
[BSA (1 mg/ml)]
[NaCl (0.5 M)]
[Adapter MseI (50 µM)]
[Adapter EcoRI (5 µM)]
[MseI (10 U/µl)]
[EcoRI (20 U/µl)]
[T4 DNA Ligase (5U/µl)]
?. [Restriction-ligation reaction]
Incubate at 37ºC for 3 h and at 17 °C overnight.
?. [Restriction-ligation reaction]
Dilute ligated DNA fragments 10-fold.
?. [Preselective PCR]
Prepare the reaction mix:
[water]
[RedTaq Buffer (10x)]
[dNTP (2.5 mM each)]
[Presel. primer Eco-A (10 µM)]
[Presel. primer Mse-C (10 µM)]
[Sigma RedTaq (1U/ µl)]
?. [Preselective PCR]
Add 2 μl of the 10x diluted restriction-ligation product to 8 μl of the reaction mix (total volume 10 μl).
?. [Preselective PCR]
Run the PCR program:72°C for 2 min25 cycles of:  94°C for 1 s
 56°C for 30 s
 72°C for 120 s
  60°C for 30 minhold at 4°C
?. [Selective PCR]
Prepare reaction mix:
[water]
[RedTaq Buffer (10x)]
[dNTP (2.5 mM each)]
[Sigma RedTaq (1U/ µl)]
[Sel. primer Mse-Cxx (5 µM)]
[Sel. primer Eco-Axx (1 µM)]
?. [Selective PCR]
Run the PCR program:94°C for 2 min10 cycles of:  94°C for 1 s
 65°C for 30 s, with a ramp of -1°C per step (up to 56°C)
 72°C for 120 s
  22 cycles of:  94°C for 1 s
 56°C for 30 s
 72°C for 120 s
  60°C for 30 minhold at 4°C
?. [Preselective PCR]
Dilute the PCR product 10-fold.
?. [Purification]
Proceed with Sephadex purification protocol and loading on ABI 3130.
?. [Restriction-ligation reaction]
Add 6 μl of DNA to 6 μl of the restriction-ligation mix.
?. [Selective PCR]
Add 2 μl of the 10x diluted preselective PCR product to 8 μl of the reaction mix (total volume 10 μl).
TITLE: Genotyping by next generation sequencing
AUTHORS: Hanqin  Li, Yogendra Verma, Dirk Hockemeyer, Frank Soldner

[DESCRIPTION]
This collection describes a standard genotyping procedure using next generation sequencing (NGS) in the Hockemeyer lab.

Collection overview
Preparing of genomic DNA from in vitro cultured cells
    A. Preparing crude cell lysate directly from hPSCs culture
    B. Preparing crude cell lysate from dissociated cells
    C. Preparing cell lysate from FACS-sorted cells
Amplifying the target genomic region by PCR
Preparing samples for NGS
CRISPResso analysis

General notes:
1. Throughout these protocols, the term hPSC is used to collectively refer to both hiPSCs and hESCs. All described procedures have been tested and work equally well for hiPSCs and hESCs.

[STEPS]
TITLE: three hours sleep restriction protocol
AUTHORS: longzhiliang 

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>This protocol was obtained from the Sleepy Brain Project (</span><a href="https://openneuro.org/datasets/ds000201" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">https://openneuro.org/datasets/ds000201</span></a><span>). The detailed protocol procedure can be found at the site. </span></div><div class = "text-block">The project features a cross-over comparison between one night sleep restriction (3 hours sleep) and normal sleep. Participants were randomized to undergo both conditions in a counterbalanced order with an interval of approximately 1 month, and completed several questionnaires such as the Epworth sleepiness scale (ESS), the Insomnia Severity Index (ISI), the Karolinska Sleep Questionnaire (KSQ) sleep quality index and snoring symptom index, and the Positive and Negative Affect Schedule (PANAS).  Among the questionnaires, the PANAS score was recorded in  both conditions of sleep restriction and normal sleep.</div></div>

[STEPS]
?. Participants were recruited by poster and newspaper advertising based on the following inclusion coriteria: 1) those required to undergo fMRI procedures, e.g., no ferromagnetic items in the body, not claustrophobic, and not pregnant; 2) those who have no current or past self-reported psychiatric or neurological illness; 3) those who do not have hypertension or diabetes; 4) those who do not use psychoactive or immune-modulating drugs; 5) those who do not consume nicotine every day.
?. Participants fill in some sleep diaries e.g. normal time in bed for three nights before the experiment.
?. On the night before MRI imaging, participants slept in their home for only three hours (sleep restriction) or as usual. Their sleep was monitored by using ambulatory polysomnography. The total sleep time during the night was recorded. In the condition of sleep restriction, they were instructed to go to bed three hours before the time they would usually wake up and get up at their normal schedule.
?. On the day of experiment, participants was told to avoid coffee, alcohol, and naps .
?. In the evening following sleep restriction or normal sleep, participants undergo MRI scanning.
TITLE: Congo Red Immunostaining
AUTHORS: daniel.dautan, Per Svenningsson

[DESCRIPTION]
Staining for amyloid structures in protein aggregates

[STEPS]
1. Wash freshly sectioned tissues 2-3 times with 1X PBS to remove OCT. 
2. Mount all sections onto microscope slides and dry at Room temperature  . 
3. Transfer slides into a 1% CongoRed solution for 30 min.
4. Place slides into alkaline bath (1% Sodium hydroxide in 50% ethanol) for 5 min. 
5. Transfer slides into 3 sequential baths of water to remove excess staining.
6. Transfer slides into 0.4% Toluidine blue solution for 10 min.

7. Wash slides in 3 sequential bath of water to remove excess staining. 
8. Finally, dehydrate slides using sequential baths of: 
distilled water (~2 min)
70% ethanol (2 times ~2 min)
95% ethanol (2 times ~2 min)
100% ethanol (2 times ~2 min)
10% xylene (2 times ~5 min) 
9. Dry sections at room temperature (~5 min).
10. Cover with DPX mounting medium.
TITLE: BSCI:414 Lab 8 Transform and Miniprep Plasmid Containing SARS-CoV-2 Spike Gene
AUTHORS: Harley King

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Received 3ul plasmid containing SARS-CoV-2 spike gene from Xiaopin Zhu at UMD. I performed transformation and plated on LB plates containing 100ug/ml ampicillin. </div><div class = "text-block">I selected a positive colony and grew overnight in 5mL LB media supplemented with 100ug/mL ampicillin. </div><div class = "text-block">A miniprep was performed on the culture.</div></div>

[STEPS]
?. Watch miniprep video.
?. Transform "pHDM-Spike" plasmid containing SARS-CoV-2 spike gene into chemically competent Dh10B cells and select on LB ampicillin plates.
?. Receive plasmid containing SARS-CoV-2 spike gene from Xiaoping Zhu @ UMD VetMed.Link to plasmid in Benchling.
?. Quantify plasmid concentration using Nanodrop Spectrophotometer.
?. Great. Purified plasmid. What can we do with it?
TITLE: Frozen stocks
AUTHORS: Elizabeth Fozo

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Preparation of frozen stocks for the PERMANENT lab collection</div></div>

[STEPS]
?. [Steps]
Begin a 10 mL overnight of the strain in an appropriate media with antibiotics as necessary. Incubate at the appropriate temperature overnight.
?. [Steps]
In the morning harvest the culture by spinning cells down at 3500 RMP for 10 minutes (program 1).
?. [Steps]
Resuspend the pellet in 1 mL of the media used to grow the strain
?. [Steps]
Add 1 mL of 80% glycerol and mix the culture well.
?. [Steps]
Transfer approximately 1.7 mL of the suspension (do not overfill the tube!) to a Nunc Cryovial
?. [Steps]
Label the tube with the organism, what it is, and any antibiotic resistance or growth specifics (if it only grows at 30oC, then say so!)
?. [Steps]
The vail must be given to the instructor with a note containing the following informationStrain backgroundWhat it is (deletion of gene X, etc.) Antibiotic resistanceTemperature restrictionsIf it is a plasmid: plasmid backbone (i.e. pBR-pLac, pLT06, etc.)Your nameThe date
TITLE: Fluorescence recovery after photobleaching (FRAP)
AUTHORS: Xinbo Wang, Pietro De Camilli

[DESCRIPTION]
This protocol details methods of the FRAP analysis of LRRK2-induced liposome tubules in vitro

[STEPS]
SECTION: Fluorescence recovery after photobleaching (FRAP)
1. Prepare LRRK2-liposome mixtures in a PCR tube with 300 nanomolar (nM) GFP-LRKK2, 20 micromolar (µM) liposomes (labeled with trace amounts of rhodamine-PE) and 1 millimolar (mM) GMPPNP.
SECTION: Fluorescence recovery after photobleaching (FRAP)
2. Immediately deposit6 µL -10 µL samples of step 1 on a   glass bottom dish and incubate at 37 °C for 30 min.
 

SECTION: Fluorescence recovery after photobleaching (FRAP)
3. Perform FRAP experiments  with a Spinning disk confocal (SDC) microscopy at Room temperature on a Nikon Ti-E inverted microscope using the Improvision UltraVIEW VoX system, with the settings as:
SECTION: Fluorescence recovery after photobleaching (FRAP)
3.1. Acquire the time-lapse images at every 15 s.
SECTION: Fluorescence recovery after photobleaching (FRAP)
3.3. Bleach three ROIs with a   laser for 500 ms.
SECTION: Fluorescence recovery after photobleaching (FRAP)
3.2. Acquire three images before bleaching.
SECTION: Fluorescence recovery after photobleaching (FRAP)
3.4. Acquire post-bleach images up to 10 min at 15 s intervals.