Suspension Cell Immunofluorescence | Shi-fix™

What is the SHI-FIX™ coating material?

It is a combination of a recombinant protein and two synthetic organic compounds.

How do suspension cells adhere to SHI-FIX™ coverslips?

The combination of hydrophilic and hydrophobic interactions between the SHI-FIX™ coating molecules and the cell membranes leads to the attachment of the cells to the coverslips.

What cell types are SHI-FIX™ coverslips validated for?

Jurkat,  K562,  U937,  Daudi,  PC12,  Raw264.7,  MOLT4, PBL culture, THP1, human PBMC,  Mouse splenocytes,  Mouse thymocytes,  Rat splenocytes,  Rat thymocytes,  HL-60,  T-ALL, NCI-H69

Does the of SHI-FIX™ coating stimulate the cells or alter their physiology?

As far as we know, SHI-FIX™ coatings does not stimulate the cells or alter their physiology apart from immobilizing them. We have also not observed any toxic effects on the cells.

What is the difference between SHI-FIX™ and SHI-FIX™ Plus coverslips?

Size:  SHI-FIX™ coverslip is 12mm in diameter, SHI-FIX™ Plus coverslip is 15mm.

 Quality: Plus line is made with high quality German glass for high resolution imaging.

 Culture maintenance: Plus line is also suitable for culturing the suspension cells for a day or two prior to staining

Application: The Plus line is better for attaching cell lines which are more difficult, e.g. K562, PBMC and other blood related cell lines

What is the difference between SB-Shi-fix96 and SB-Shi-fix96Plus well plate?

SB-Shifix96 is for cell attachment followed by staining and imaging.

SB-Shifix96Plus allows you to maintain the attached cells in culture for up to 48 hours before staining.

Is it possible to do 3D culture of the cells in SHI-FIX™ coverslips?

This has not been tested.

Is there use of any human origin material to produce SHI-FIX™ products?

No

What is the shelf life of the SHI-FIX™ and SHI-FIX™ Plus coverslips? 

The shelf life for both SHI-FIX™ and SHI-FIX™ plus is one year.

How do I store SHI-FIX™ products ?

Recommended storage condition is 4^oC to 8^oC.

How long can cells attached to SHI-FIX™ be cultured continuously?

Cells can be maintained on Shi-fix PLUS coverslips or plates for up to 5 days in culture.

Is it possible to fix microorganisms, free mitochondria, organelle membranes, and mouse embryos (with transparent body)?

Yes, it is possible to fix microorganisms like E.Coli and organelle membranes. However, mouse embryos have not yet been tried.

Is it possible to measure the metabolic activity of the attached cells?

Yes

Can the cells bound to Shi-fix can be peeled off?

This has not been tried.

How much of the cell population is lost after 3 washes?

A loss of 10-20% of total cells (depending on the cell types) is expected after 3 washes.

Is it possible to attach a mixture of adherent and suspension cells to the coverslips?

Yes. Add adherent cells to the coverslips first, wait for 30 min, then add  suspension cells and wait for another 30 min before washing.

If the number of cells on the coverslips is insufficient after adding the cell suspension, can you can add the cell suspension again?

Yes

Do the cells adhere to the coverslips in a double layer ?

No. The attachment is a monolayer.

Can the Shi-Fix coverslips be applied to the study of CAR-T immune synapses?

We haven't tested yet however it is probably possible to use Shi-fix coverslips to the study of CAR-T immune synapses. Recently,  we co-cultured adherent and suspension cells (K562 and HeLa ) on the SHI-FIX™ coverslips and the cells were very healthy and growing in a monolayer (K562 cells and HeLa cells co cultured).

Have you tested the lower limit of applicable cells on SHI-FIX™ COVERSLIPS? 

we have used a minimum 15000 cells per coverslip. Cells fewer than that probably need to be resuspended in a very small volume- 10-20 ul.

Are SHI-FIX ™ COVERSLIPS and Plates all sterile? 

Yes, all our SHI-FIX™ products are sterile.

Can we seed cells without washing with PBS?

Yes, you can seed cells without washing with PBS but the attachment of cells will be very low.

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