Apr 17, 2025

Public workspaceNRP1_01_ImagePlate_Coating V.2

This protocol is a draft, published without a DOI.
  • 1Washington University in St. Louis;
  • 2Washington University School of Medicine @FIVE;
  • 3Washington University School of Medicine;
  • 4Five MGI;
  • 5Washington University: MGI;
  • 6Washu
William J Buchser
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Protocol Citation: William J Buchser, Jason Waligorski, Colin Kremitzki, Lillian Gaddy, Emanuel Gerbi, Caleb Valentine 2025. NRP1_01_ImagePlate_Coating. protocols.io https://protocols.io/view/nrp1-01-imageplate-coating-eajgbacjxVersion created by William J Buchser
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: April 10, 2025
Last Modified: April 17, 2025
Protocol Integer ID: 129352
Disclaimer
MGI at Washington University School of Medicine in St Louis
Abstract
NRP Milestone # 1 Plate Coating Protocol
Plating Schedule
Plating Schedule
Locate weekly plating schedule in FMD to determine how many 384 and 96 well plates are needed.
FIVE Master Data "NRP PlateTracker"

Copy the Template Folder Structure Template and Rename it for the PlateID being worked on
Template Folder: R:\FIVE\EXP\FIW103\P999\
Contents of Template Folder

Ensure that enough media is stocked for the amount of plating that will be done
Plating Layout and Labels
Plating Layout and Labels
Print labels on the Brady Label Printer.
Label information should include:
  • FIW###P##
Large label should be placed on bottom right corner of 384 or 96 well plate lid.
Small labels should be placed on right corners of the actual plates.

384 Well Example of Label Placement

iMatrix Dilution
iMatrix Dilution
iMatrix-511 Silk (892 021) stored at 4C. Found in the mini iPSC refrigerator.
Record Lot # of iMatrix found on side of box.
Prepare diluted stocks of iMatrix for corresponding plates by diluting in DPBS -/-.
Step case

96 Well iMatrix Coating
7 steps

iMatrix dilution factors:
  • 96-wells= 1:100
1200ul of a 1:100 dilution is needed for a single 96 well Ab plate.
Prepare in a 15mL conical.
For a 1:100 dilution you will need 1200ul of DPBS -/- to 12ul iMatrix.
Manually dispense 100ul of 1:100 diluted iMatrix stock per well into 96 well Revvity 96 well optical plate using Integra 1250 pipette on repeat.
Wells B2-3,C2-3,D2-3,E2-3.
96 well plate incubated at 37C,5% CO2 for 1hr prior to cell seeding.