eamag/protocols-with-steps huggingface dataset:
TITLE: Formation of gp37 LTF needle trimers-LPS complex
AUTHORS: Mohammad Z. Islam, Andrei Fokine, Marthandan Mahalingam, Zhihong Zhang, Carmela Garcia-Doval, Mark J. van Raaij, Michael G. Rossmann, and Venigalla B. Rao

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Tailed bacteriophages (phages) are one of the most abundant life forms on Earth. They encode highly efficient molecular machines to infect bacteria, but the initial interactions between a phage and a bacterium that then lead to irreversible virus attachment and infection are poorly understood. This information is critically needed to engineer machines with novel host specificities in order to combat antibiotic resistance, a major threat to global health today. The tailed phage T4 encodes a specialized device for this purpose, the long tail fiber (LTF), which allows the virus to move on the bacterial surface and find a suitable site for infection. Consequently, the infection efficiency of phage T4 is one of the highest, reaching the theoretical value of 1. Although the atomic structure of the tip of the LTF has been determined, its functional architecture and how interactions with two structurally very different Escherichia coli receptor molecules, lipopolysaccharide (LPS) and outer membrane protein C (OmpC), contribute to virus movement remained unknown. Here, by developing direct receptor binding assays, extensive mutational and biochemical analyses, and structural modeling, we discovered that the ball-shaped tip of the LTF, a trimer of gene product 37, consists of three sets of symmetrically alternating binding sites for LPS and/or OmpC. Our studies implicate reversible and dynamic interactions between these sites and the receptors. We speculate that the LTF might function as a “molecular pivot” allowing the virus to “walk” on the bacterium by adjusting the angle or position of interaction of the six LTFs attached to the six-fold symmetric baseplate.</div></div>

[STEPS]
TITLE: LRRK2 Immunofluorescent staining 
AUTHORS: sherbst  

[DESCRIPTION]
Protocol for immunofluorescent staining for LRRK2 in cultured cells using the MJFF2 (c41-2) antibody. 

[STEPS]
1. Culture cells as usual on cover slips or in a plate suitable for imaging.

If using coverslips, we recommend Ø13 mm for cells cultured in 24-well plates.  
 

2. Fix cells in 4 % PFA/PBS at 4 °C for 15 min.  

4. Permeabilise the plasma membrane by incubating samples in ice-cold MeOH for 10 min. This can be done on the bench or the whole plate can be put into the -20 °C freezer. Make sure that samples are always submerged. 

3. Optional: at this step, coverslips or plates can be stored at 4 °C in PBS. Never allow samples to try out. 
5. Wash samples once in PBS
6. Incubate samples in blocking buffer for 20 min at Room temperature 

7. Incubate samples in primary antibody solution ( MJFF2 (c41-2) antibody diluted 1:100 in blocking buffer) for 60 min at Room temperature 
 



8. Wash samples three times in PBS

9. Incubate samples in secondary antibody solution ( eg anti-rabbit-Alexa Fluor™ 488 diluted 1:800 in blocking buffer) for 45 min at Room temperature  in the dark
 



10. Wash samples twice in PBS
11. Incubate sample in DAPI staining solution for 10 min at Room temperature  in the dark

12. Wash samples twice in PBS
13. Mount coverslips onto slides using a mounting medium of choice (eg DAKO Fluorescence Mounting medium) and let try at RT in the dark.
If using well-plates, add PBS to plates. 
Keep samples in the dark and store them at 4 °C for short-term storage.
TITLE: DNA extraction for human microbe samples.
AUTHORS: Lilan Hao

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol is used to clarity the process of total DNA extration for human microbe samples.</div></div>

[STEPS]
?. 200μl samples were mixed with 500 μl TE buffer and 20 μl of 10 mg/ml lysozyme solution.
?. Incubate at 37 °C for 90min. Invert the tube several times to mix every 15 minutes. Centrifuge at 13,000 rcf/min for 10 min at 4 °C and discard the supernatant.
?. Add 300 μl of 20 mg/ml proteinase K and 5μl of 10% SDS. Vortex to resuspend cells.
?. Incubate at 55 °C in a shaking water bath for 120 min to effect complete lysis. Invert the tube several times to mix every 15 minutes.
?. Centrifuge at 13,000 rcf/min for 10 min at 4 °C, then transfer the supernatant to a nuclease-free 2 ml microfuge tube.
?. Add 700 μl phenol-chloroform-isoamylalcohol (25:24:1) and vortex to mix well, centrifuge at 13,000 rcf/min for 10 min at 4 °C, then transfer the supernatant to a nuclease-free 2 ml microfuge tube.
?. Add 700 μl chloroform-isoamylalcohol (24:1) and vortex to mix well. Centrifuge at 13,000 rcf/min for 10 min at 4 °C, then transfer the supernatant to a nuclease-free 1.5 ml microfuge tube.
?. Add 1/10 volume of 3M sodium acetate, 2 μl of 5 mg/ml glycogen and 800 μl cold isopropanol. Incubate at -20 °C for the night.
?. Centrifuge at 13,000 rcf/min for 10 min at 4 °C, then discard the supernatant.
?. Washed with 800 μl of 70% ethanol. Centrifuge at 13,000 rcf/min for 10 min at 4 °C, then discard the supernatant.
?. Wash the precipitate with a second 800 μl of 70% ethanol and centrifuge as above. discard the supernatant.
?. Let sit for 15 min to dry the precipitate. Add 1×TE buffer to resuspend the precipitate.
?. Add 1μl RNase A to samples and invert tube several times to mix. Incubate at 37 °C for 30 minutes.
TITLE: Handling of genomics samples
AUTHORS: Adriana  Alberti, Julie Poulain, Stefan Engelen, Karine Labadie, Sarah Romac, Isabel Ferrera, Guillaume Albini, Jean-Marc Aury, Caroline Belser, Alexis Bertrand, Corinne Cruaud, Corinne Da Silva, Carole Dossat, Frédéric Gavory, Shahinaz Gas, Julie Guy, Maud Haquelle, E'krame Jacoby, Olivier Jaillon, Arnaud Lemainque, Eric Pelletier, Gaëlle Samson, Marc Wessner, Genoscope Technical Team, Silvia G. Acinas, Marta Royo-Llonch, Francisco M. Cornejo-Castillo, Ramiro Logares, Beatriz Fernández-Gómez, Chris Bowler, Guy Cochrane, Clara Amid, Petra Ten Hoopen, Colomban De Vargas, Nigel Grimsley, Elodie Desgranges, Stefanie Kandels-Lewis, Hiroyuki Ogata, Nicole Poulton, Michael E. Sieracki, Ramunas Stepanauskas, Matthew B. Sullivan, Jennifer R. Brum, Melissa B. Duhaime, Bonnie T. Poulos, Bonnie L. Hurwitz, Stéphane Pesant, Eric Karsenti, Patrick Wincker

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>This protocol describes the handling of genomics samples for the </span><span style = "font-style:italic;">Tara</span><span> Oceans expedition and is part of </span><a href="https://www.protocols.io/view/viral-to-metazoan-marine-plankton-nucleotide-seque-qv6dw9e" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">Viral to metazoan marine plankton nucleotide sequences from the </span><span style = ":;font-style:italic;">Tara</span><span style = ":;"> Oceans expedition</span></a><span>. </span></div><div class = "text-block"><div class = "justify" style = "text-align:center"></div></div><div style = "text-align :; float : ;"><img style = "" src = "https://s3.amazonaws.com/pr-journal/zhnfw26.jpg" /></div><div class = "text-block"><div class = "justify" style = "text-align:left"><span style = "font-weight:bold;">Figure 1</span><span>: Overview of -omics analysis strategy applied on Tara Oceans samples.</span></div></div></div>

[STEPS]
TITLE: 6: Protocol optimization for SABER-FISH in tissues
AUTHORS: Jocelyn Y. Kishi, Sylvain W. Lapan, Brian J Beliveau, Emma R. West, Allen Zhu, Hiroshi M. Sasaki, Sinem Saka, Yu Wang, Constance L Cepko, Peng Yin

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">This protocol summarizes several tested variations to simplify the SABER-FISH protocol in tissues; In most cases robust signal was found.</div></div>

[STEPS]
?.
TITLE: Glycerol stocks 
AUTHORS: iGEM Dusseldorf

[STEPS]
?. Overnight liquid culture (3mL) + Antibiotic.For E.coli (Low salt) LBFor Pichia YPD
?. 80% Glycerol: mix 20mL H2O with 80mL pure glycerol and then autoclave
?. E. coli: 812µL of cells mix with 188 µL of sterile 80% glycerol
?. Pichia: Spin 2min at 2000g. Decant 2ml. Resuspend in remaining liquid. 812µL of cells mix with 188µL of sterile 80% glycerol
?. Use cryovials for freezing away. Store at -80° C
TITLE: WU sc-prep Protocol for Solid Tumors v2.1
AUTHORS: Reyka Jayasinghe, Li Ding, Feng Chen

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block">Single-cell Isolation for solid tumors  Ding Lab 2021</div></div>

[STEPS]
?. [Reagents]
-Human tumor dissociation kit(Miltenyi Biotec #130-095-929)-PBS-PBS, 0.5% BSA (keep at 4°C)-(optional) Red Blood Cell (RBC) Lysis Solution ACK buffer Fisher (A1049201)-Falcon 15mL/50mL Conical Centrifuge Tubes (Corning Life Sciences DL, Fisher Scientific)-Large filter: Fisherbrand Cell Strainers. Nylon mesh 40um (Fischer Scientific, Cat # 22-363-547)-Smaller filter: PluriSelect Mini-Strainer 40 uM (Cat #43-10040-60)-Petri dishes -LoBind Protein or Genomic Microcentrifuge Tubes, Eppendorf®, Volume=2 ml Fisher 13-698-792-All steps should be performed under sterile conditions-For tissue fragments >1000mg, scale up reagent volumes (volumes listed are for 1 g tissue)
?. [Octo Dissociation]
Mince tissue into small pieces with razor blade in petri dish.
?. [Octo Dissociation]
Resuspend tissue in enzyme mix (2.5 mL DMEM to 100 uL of Enzyme H, 50 uL Enzyme R, and 12.5 uL enzyme A) in C-Tube
?. [Octo Dissociation]
Incubate on gentleMACS™ Octo Dissociator -  37h_TDK_1 setting (incubation can be shortened if tissue is very soft ~20-25 min). Check sample every 10-15 minutes.
?. [Octo Dissociation]
After dissociation, filter cells through 40um filter and spin down in 15ml tube at 450g for 5 min at 4 degrees
?. [Octo Dissociation]
If there is any red visible in the cell pellet proceed to ACK RBC lysis step below (Refer to bottom of protocol for examples).RBC ACK Lysis Solution-Slowly resuspend cell pellet in 500 uL-5mL (5ml for larger pellets) of RBC ACK Lysis Solution-Incubate on ice for 30 sec to 5 min (longer incubation for more RBCs)-Quench reaction with 10mL of BSA/PBS buffer-Centrifuge at 450g 5min at 4 degrees
?. [Octo Dissociation]
Do 1-3 1mL washes of 0.5% BSA in PBS on ice (spin at 450g 5min at 4 degrees) to remove dead cells and debris. Do washes in 2mL tube. After each wash aspirate off BSA/PBS and leave ~50-100 uL of BSA/PBS buffer. Resuspend cells using 1000 uL wide-bore pipette tip and count 10 uL of sample using a hematocytometer and trypan blue.After first wash take 10 uL of sample and visualize under microscope (for smaller amounts take 2 uL of sample and 8 uL of wash buffer). If after 1 wash there are too many dead cells – consider using dead cell removal kit or debris removal kit: Dead Cell Removal: https://www.miltenyibiotec.com/US-en/products/macs-sample-preparation/removal-reagents/dead-cell-removal-kit.htmlDebris Removal: https://www.miltenyibiotec.com/US-en/products/macs-sample-preparation/removal-reagents/debris-removal-solution.htmloNote: For Debris removal – use slow speed 450-500 g for all spinning steps.If cell suspension looks pretty clean after first wash, then second wash should be clean to load. oIf you have approximately 400-2000 cells/uL then continue on to GEMs if doing single cell. Load 20,000 cells.
TITLE: Live-cell imaging; reactive oxygen species (Superoxide)
AUTHORS: Minee-Liane Choi

[DESCRIPTION]
This protocol contains the instruction for measuring superoxide using dihydroethidium (HEt) which allows the rate of superoxide generation to be measured which is present as the ratio of the oxidized form of the dye over the reduced form. HEt allows the rate of cytosolic superoxide generation to be measured, by the slope of the ratio of oxidized dye to reduced dye.

[STEPS]
1. Cells are washed and loaded 2 micromolar (µM) dihydroethidium (HEt; Thermo Fisher Scientific) in the recording buffer. 
 
 


2. The recording is performed using an epi-fluorescence inverted microscope equipped with 20x objective after a quick loading (2-3 min) in order to limit the intracellular accumulation of oxidized product, and the dye was present throughout the imaging. 
 

3. Excitation is at530 nm, and emission recorded above 560 nm is assigned to the oxidized form, while excitation at 380 nm and emission collected from 405 nm to 470 nm is assigned to the reduced form. 
4. The ratio of the fluorescence intensity, resulting from its oxidized/reduced forms, is quantified.
TITLE: Cleaning an OT-2 COVID-19 Diagnostic Station
AUTHORS: Max Marrone

[STEPS]
?. Wipe these parts of the OT-2 down with a 1:10 dilution of bleach:The clear polycarbonate windows.The black pipette stems. (Avoid the rest of the pipettes, including the ejectors.)The aluminum deck.The removable black trash bin.
?. Wait , then quickly rinse the bleach off with distilled water.
The aluminum on the OT-2 will be discolored if the bleach sits for too long. In the long term, it may also cause more serious corrosion.
?. Wipe these parts of the OT-2 down with RNaseZap or RNase AWAY.The same parts that you wiped down with bleach:The clear polycarbonate windows.The black pipette stems. (Avoid the rest of the pipettes, including the ejectors.)The aluminum deck.The removable black trash bin.Plus these additional parts:The bottoms of the pipette ejectors.Any Temperature Modules or Magnetic Modules that the OT-2 has on its deck.Any 96 well aluminum blocks that are going to be used on the OT-2.Plus these additional parts:The bottoms of the pipette ejectors.Any Temperature Modules or Magnetic Modules that the OT-2 has on its deck.Any 96 well aluminum blocks that are going to be used on the OT-2.
?. Rinse the RNaseZap or RNase AWAY off with distilled water.
?. Wipe the OT-2 dry, or let the water evaporate.
TITLE: No Title
AUTHORS: 

[GUIDELINES]
Materials:<br />
<p>1) <em>Chlorella</em> virus, in 50 mM Tris-HCl, pH 7.8</p>
<p>2) 100 mM Tris-HCl, pH 7.4, 10 mM EDTA, 1 M NaCl (10X TEN, pH 7.4)</p>
<p>3) 1.0% Na sarcosyl</p>
<p>4) 10 mM Tris-HCl, pH 8.0, 1 mM EDTA (1X TE)</p>
<p>5) Isopropanol, saturated with CsCl/TE buffer</p>
<p>6) 3 M NaOAc</p>
<p>7)  Hoechst dye #33258, 100 µg/mL<br />8) CsCl gradients: 40-60% (w/w) CsCl gradients made up in SW60 rotor ultra clear tubes, equilibrated with 1X TE, pH 8.0 + 1 µg/mL Hoechst dye (see recipe)<br /><br /></p>
<table>
<tbody>
<tr>
<td>%</td>
<td>gm CsCl</td>
<td>ml 1X TE</td>
<td>µl Hoechst dye <br />(100 µg/mL)</td>
<td>layer (µl)</td>
</tr>
<tr>
<td>60</td>
<td>9.0</td>
<td>6.0</td>
<td>60.0</td>
<td>1000</td>
</tr>
<tr>
<td>50</td>
<td>7.5</td>
<td>7.5</td>
<td>75.0</td>
<td>1050</td>
</tr>
<tr>
<td>40</td>
<td>5.0</td>
<td>7.5</td>
<td>75.0</td>
<td>1050</td>
</tr>
</tbody>
</table>
<br />9) 60% CsCl (w/w): 9.0 gm CsCl, 6.0 ml 1X TE, pH 8.0, 120.0 µL Hoechst dye (100 µg/mL)

[STEPS]
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?. 
?.
TITLE: Glufosinate ammonium (BASTA) Paint Assay for Screening for BAR Resistance Gene in Tobacco
AUTHORS: Lynn  Doran

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Glufosinate ammonium (BASTA</span><span style = "vertical-align:super;">TM</span><span>/LIBERTY</span><span style = "vertical-align:super;">TM</span><span>) Paint Assay for Screening for the presence of the </span><a href="https://benchling.com/ripe/f/lib_UNvrVcvh-registry/seq_7ktFxC9l-bar/edit" style = "text-decoration:underline;color:blue;cursor:pointer;"><span style = ":;">BAR Resistance Gene</span></a><span> in Transgenic Tobacco.</span></div></div>

[STEPS]
?. [Herbicide Dilution]
Make a working stock of 1.5 g/L of glufosinate ammonium.
?. [Herbicide Dilution]
Determine the amount of glufosinate ammonium active ingredient  by reading the EPA herbicide label.Liberty 280 SL contains 280 g/L.
?. [Herbicide Dilution]
Calculate and perform the required dilution from the amount of active ingredient in the stock herbicide to obtain a working stock of 1.5 g/L. Liberty 280 SL (280 g/L) will be diluted 160 ul Liberty 280 SL into 30 mL of distilled water.
Wear appropriate PPE as prescribed by the herbicide label at all times when handling the herbicide.  Notify workers around you that you will be applying herbicide.  Ensure that all workers who will be working in areas where herbicide is applied have been trained annually on the EPA Worker Protection Standard.
?. [Herbicide Application]
Draw a line across the upper surface of a healthy leaf with a permanent marker.
?. [Herbicide Application]
Apply the working stock (1.5 g/L glufosinate ammonium) herbicide to the section of the leaf from the tip to the permanent marker line.
?. [Paint Assay Interpretation]
After approximately 5 days, observe the leaf.  If the leaf has died to approximately the level of the marker line, the plant is glufosinate ammonium sensitive indicating that it most likely does not contain the BAR resistance gene.  If the leaf has died entirely or well past the marker line, the plant may be experiencing physiological distress (water stress, disease pressure, etc) unrelated to the paint assay.  The assay is inconclusive.  Correct the growth conditions to remove the plant stressor and repeat the assay on a healthy leaf.  If the leaf does not exhibit necrosis on the leaf from the tip to the marker line, the plant most likely contains the BAR resistance gene.If the leaf has died entirely or well past the marker line, the plant may be experiencing physiological distress (water stress, disease pressure, etc) unrelated to the paint assay.  The assay is inconclusive.  Correct the growth conditions to remove the plant stressor and repeat the assay on a healthy leaf.  If the leaf does not exhibit necrosis on the leaf from the tip to the marker line, the plant most likely contains the BAR resistance gene.
TITLE: Phage infection and timed harvest of E. coli and B. subtilis cells 
AUTHORS: Januka  Athukoralage, Adair  Borges

[DESCRIPTION]
This protocol outlines how to infect E. coli and B. subtilis with phage T4 and SPO1, respectively, and harvest cells at set time points.

Harvesting cells at different timepoints during infection allows you to evaluate when bacteriophage transcripts are most abundant in cells and maximize their enrichment for downstream analyses such as chemical analysis and RNA sequencing.

[STEPS]
SECTION: Bacteriophage infection
1. Prepare 30 mL subcultures of E. coli and B. subtilis at 1:100 dilution (300 µL in 30 mL) and grow to a target OD600 of 0.4 in a shaking incubator (180 rpm). For both E. coli and B. subtilis this is ~2 h at 37 °C. Supplement the subculture media (LB) with 1 mM CaCl2 and 1 mM MgCl2.
SECTION: Bacteriophage infection
2. Aliquot 500 µL of cell culture into several 1.5 mL microcentrifuge tubes and inoculate with 50 µL of phage at the correct dilution to reach your target MOI (see our protocol on MOI calculations, linked below).
 

SECTION: Cell harvest
5. Pre-cool a bench-top centrifuge to 4 °C and at desired time points, harvest cells by centrifuging at 5000 × g for 3 min. For example, cells can be harvested at 5 min and 15 min in the hope of capturing both earlier and later infection stages. This timing will be highly phage-dependent! 
SECTION: Cell harvest
6. Remove the supernatant from the pellet and immediately flash-freeze the pellets in liquid nitrogen. You must be speedy during this process to effectively halt phage infection. You can store bacterial pellets at –80 °C and use later for downstream applications such as RNA extraction. 
SECTION: Bacteriophage infection
4. Plate out phage and bacteria to calculate the actual MOI used in the experiment.

SECTION: Bacteriophage infection
3. Here, we are infecting E. coli with T4 and and B. subtilis with SPO1.  Be sure to simultaneously carry out a control experiment where you mock-infect 500 µL E. coli and B. subtilis cultures with 50 µL sterile SM buffer. Once inoculated, incubate the microcentrifuge tubes at 37 °C on a heated shaker block, shaking at 300 rpm.
TITLE: Creating Plate Layout in FIVTools
AUTHORS: Jason Waligorski

[DESCRIPTION]
Plate layout creator in FIVtools

[STEPS]
SECTION: Creating Plate Layout in FIVTools
1. Open FIVTools
SECTION: Creating Plate Layout in FIVTools
2. Click Layouts


 


SECTION: Creating Plate Layout in FIVTools
3. Update Exp Name field (highlighted in blue above) to the corresponding FIV experiment Number (make sure to keep “FIV” in the name)
SECTION: Creating Plate Layout in FIVTools
4. Click once on the Export Folder field so it updates the folder location based on the corresponding FIV experiment number
SECTION: Creating Plate Layout in FIVTools
5. Choose the desired number of 96-well plates by typing 1, 2, 3, or 4 in Count


 


SECTION: Creating Plate Layout in FIVTools
6. Select 96 Mote to exclude plating from the outermost wells

 


SECTION: Creating Plate Layout in FIVTools
7. Select "384 Full" for a 384 well plate
SECTION: Creating Plate Layout in FIVTools
8. If the layout is not standard (does not fall within 96 mote, 96 full, or 384 full), adjust the First Row, First Column, Last Row, and Last Column fields to fit your custom plate format. 


 


SECTION: Creating Plate Layout in FIVTools
9. If doing an antibody plate, check the “Ab Plate” box and:
Make “Count” = 0 for one antibody plate
Make “Count” = 2 for two 96-well plates and an antibody plate
SECTION: Creating Plate Layout in FIVTools
10. Change “Cells per Well” to desired plating density

 

SECTION: Creating Plate Layout in FIVTools
11. Copy experimental condition names (often genotypes, dosages, G1/G2, etc.) and click “PASTE Names” to auto-fill the 12-well source plate. 


 

SECTION: Creating Plate Layout in FIVTools
12. Name can also be manually filled
SECTION: Creating Plate Layout in FIVTools
13. The Well column can only have row/column combinations of a 12-well plate (A-C, 1-4). Please note which names (experimental conditions) correspond to which wells. 
SECTION: Creating Plate Layout in FIVTools
14. Relative Representation allows you to assign different weights for representation in the 96-well plates
SECTION: Creating Plate Layout in FIVTools
15. Use the underscore symbol to have a cell type only show up in the antibody plate layout (for example: “_cell type”)
SECTION: Creating Plate Layout in FIVTools
16. Click Fill Plates (in red)

 

SECTION: Creating Plate Layout in FIVTools
17. A randomized plate layout will be populated based on your input criteria. Each condition will have a unique color in the layout window. Try to make sure the empty wells (positions with white space/no circles) are not on any edges. Also if doing multiple plates, make sure the empty wells are oriented differently across plates. 
SECTION: Creating Plate Layout in FIVTools
18. Go into the “4 Mapping” folder of the respective FIV experiment folder and make sure there is a PlateMap, PlatingSetup, and script. 


 

SECTION: Creating Plate Layout in FIVTools
19. After source and destination plates are physically created, excute Biomek plating in accordance with 

Biomek 96-well plating - Google Docs
TITLE: Immunostaining infiltrating spheroids as preparation for quantitative light-sheet imaging
AUTHORS: Benedicte Bjørknes, Oliver Emil Neye, Petra Hamerlik, Liselotte Jauffred

[DESCRIPTION]
Although various in vivo and in vitro models for studying glioblastoma cell invasion has progressed the field, there is still a need for optimized procedures. In particular to reveal key features of glioblastoma biology and infiltrating growth. In this protocol, we present an approach using indirect immunofluorescence in a 3D human xenograft glioblastoma spheroid model embedded in a naturally derived extracellular matrix

[STEPS]