Aug 21, 2024

Public workspaceConA Cover Slide Preparation

DOI
dx.doi.org/10.17504/protocols.io.kxygxy3rzl8j/v1
  • 1UMass Chan Medical School, RNA Therapeutics Institute, Worcester, MA, USA;
  • 2University of Alberta, Department of Cell Biology, Edmonton, AB, Canada;
  • 3University of California, Department of Viticulture and Enology, Davis, CA, USA
Mathias Hammer
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DOI: dx.doi.org/10.17504/protocols.io.kxygxy3rzl8j/v1
Protocol Citation: Mathias Hammer, Ammeret Rossouw, Azra Lari, Ben Montpetit, David Grunwald 2024. ConA Cover Slide Preparation. protocols.io https://dx.doi.org/10.17504/protocols.io.kxygxy3rzl8j/v1
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: Working
We use this protocol and it's working
Created: May 28, 2024
Last Modified: August 21, 2024
Protocol Integer ID: 100780
Keywords: yeast fluorescence imaging, ConA, Concanavalin A, CanA slide, yeast fixation
Funders Acknowledgement:
NSF
Grant ID: 1917206
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Abstract
Concanavalin A is used to attach yeast cell to surfaces. This protocol describes the preparation of cover slides for high resolution fluorescence microscopy.
Materials
Concanavalin A (ConA) from Canavalia ensiformis:
Sigma-Aldrich
Cat#: C2272-10MG
Lot#: 129H0322

UltraPure Distilled Water

Cover slides ⌀ 25mm, No. 1.5 or 1.5H

Attofluor Cell Chamber
Thermo Fisher Scientific
Cat#: A7816

Equipment:

10 ml centrifugation tube
10 ml pipette

Quorum Emitech K100X glow discharger
mixer
Before start
Have the following solutions premixed:

Concanavalin A (ConA) solution:
Concentration: 1 g/l
resolve 10 mg ConA in 10 ml distilled water, vertex
store at Temperature-20 °C

Cover slide preconditioning
Cover slide preconditioning
Rinse a 25mm cover slide No. 1.5 or No. 1.5H.
Dry with Kim wipe, remove leftover driplet with optical tissue.

Plasma clean the cover slide to a hydrophilic negative charged surface.
Quorum Emitech K100X glow discharger at Amount25 mA Duration00:00:45
45s
Place the cover slide in the cell chamber.
ConA slide preparation
ConA slide preparation
30m
30m
Defrost the stock ConA of a concentration of Concentration1 mg/mL

Pipette Amount1 mL ConA solution into the cell chamber.

Let the ConA settle, covered with Kim wipe for Duration00:10:00 at the bench.

10m
Remove the ConA from the chamber back in its container.
Let the cover slip dry, covered with Kim wipe for Duration00:20:00 .
Note
The prepared ConA cover slides can be used within a week, if stored in ways that secure its not exposed to dirt and dust.


20m