coverslips for in situ culture

[Maria Mouriki]

Dear all,

Would you please share with me any tips on the coverslips you are
using on in situ cultures? Are these coverlips treated in a way/
covered with proteins that promote cells adherence? Could you suggest
a brand/manufacturer to me? Would simple coverslips do following
sterilization? I intend to use a 22x22mm c/slip in a small petri dish
for each culture (amniotic fluid, CVS and products of conception).

Thank you all in advance!
Maria

[Gwenevere Carter]

We found a company that sells sterile 22x22 coverslips in petri dishes.  The company is

MatTek Corp.

Product # is CSGK/F

1-800-634-9018

www.glassbottomdishes.com

email imfor...@mattek.com

Our one complaint is that some of the coverslips are broken in the dishes,  they have a more expensive version with form to prevent breakage.

 

Gwenevere Carter, MT(ASCP),CG(ASCP)

Cytogenetics Department

Woman's Hospital

Baton Rouge, La

 

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[laevsky ilana]

Hi Maria,

 

We are using Menzel-glaser's 22x22 coverslips for the same purpose. We are just sterilizing it in big glass petri dish (gas sterilization or regular dry autoclave). We using Nunc's small petri dishes. We don't treating c/slips for adherence, but we seeding our cells in very small amount of medium (0.5 ml) and after 24 hours adding a regular amount of medium (2ml), it's solve an adherence problem.

Regards

Ilana

From: Maria Mouriki <mend...@yahoo.com>
To: Cytogenetics methods and trouble-shooting Forum <cytogenetics-methods...@googlegroups.com>
Sent: Thursday, February 9, 2012 1:13 PM
Subject: coverslips for in situ culture

[maria mouriki]

Thank you very much Gwenevere. I will look into it and if I have any queries I will let you know.

Maria

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[maria mouriki]

Thank you very much Ilana! I will keep your suggestion in mind.

Maria

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[Gwenevere Carter]

anytime

[karyotyper]

Hi Ilana,

can I ask you which mounting medium are You using or You can recommend
to mount coverslips on slides ???

I'd like to try in situ cultivation, but I didnt find any protocol for
mounting coverslips from in situ on microscope slides. I mean mount
coverslips cell side-up. Is it possible such mounted coverslips to age
in oven at 95°C and stain them or it is necessary to age and stain
them before mounting ???

Thank You very much
Karyotyper

[laevsky ilana]

Good morning, karyotyper!

We using Merk's Entallan for mounting to slides and for closing slides after staining. According to our protocol, we mount the coverslips on slides as a last step of harvesting, than oven at 55°C overnight and stain. After staining we are air drying (or oven at 55°C for 1 hour) the slides and closing them with additional coverslips. Very simply and working good.

Good luck!

Ilana

 

From: karyotyper <petr....@centrum.cz>
To: Cytogenetics methods and trouble-shooting Forum <cytogenetics-methods...@googlegroups.com>

[Davis, Sue D.]

Hi Karyotyper,

 

In our lab we use Cytoseal 60 from Richard Allan Scientific, subsidiary of Thermo Fisher Scientific, ref # 8310-16.  We band our slides after they have aged on a hotplate at 75-80°C for approximately 2 hours and then mount them on a regular slide cell side down to avoid scratching or smudging from handling during scope analysis.  We mount them cell side up only if they are being used for FISH.  The slides can be used for analysis after the mounting media spreads completely but do not wipe oil off of the slides for at least 24 hours after they are mounted or you can easily remove the coverslip from the slide.  Good luck!

 

Susan Davis

Team Leader

Cytogenetics Laboratory

Cedars Sinai Medical Center

(310) 423-4987

 

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[karyotyper]

Thank You Ilana and Susan, for good ideas how to mount coverslips.

Best Regards

Karyotyper