X-ZELL Cryoimmunostaining™

A NEW LEVEL OF INSIGHT

X-ZELL Cryoimmunostaining™ is a patented three-step process to examine up to eight biomarkers on the single cell level without digital image processing.

It is based on a series of ground-breaking bench top instruments designed and manufactured to the highest quality by X-ZELL.

A true end-to-end solution, it covers the complete workflow of multiplexed immunostaining on standard laboratory slides – from cytocentrifugation and fixation through to immunostaining, nuclear staining and sample preservation.

Applications include rare-cell imaging, slide-based cytometry, cell-cell interaction studies and more.

All X-ZELL technologies require commercially available, fluorophore-conjugated flow-cytometry antibodies.

 

KEY ADVANTAGES:

  • Preservation of cell morphology
  • No cross-linking of antigens
  • Re-staining possible
  • Abrogation of non-specific antibody binding
  • No spectral unmixing required
  • No compensation required
  • Minimal photo bleaching
  • Ability to isolate RNA after staining (> 50% RNA preservation)
  • Full workflow in less than 3 hours with less than 45 minutes hands-on time.

Step 1: Cytocentrifugation™

Cells are spun onto gelatinized standard laboratory slides in Cell Concentrators of a StatSpin Cytofuge 2 or 12. The combination of
X-ZELL’s Cytocentrifugation™ buffer and gelatinized slides guarantees that over 90% of cells on the slides are recovered. The reagent also prevents cell dehydration, which may impact cell morphology and antigen recovery during handling after cytocentrifugation.

Time including sample preparation: 20 minutes. Hands-on time: 10 minutes.

 

MATERIALS & REAGENTS:

  • X-ZELL Cytocentrifugation™ Buffer
  • Gelatin-coated slides
  • Cell concentrators and backing plates (Beckman, USA)
  • StatSpin Cytofuge 2 or 12 (Beckman, USA)

Step 2: Cryofixation™

Cryofixation™ is a simple, highly efficient and non-toxic cell fixation process based on X-ZELL’s patented Cryofixation Station™.

In this semi-automated device, cells immobilized on slides are incubated in X-ZELL slide cartridges containing proprietary fixation and re-hydration buffers in a strictly controlled, ultra low-temperature environment – ensuring optimal preparation for immunostaining without loss of antigen and morphology.

Total incubation time: 55 minutes. Hands-on time: 3 minutes.

 

MATERIALS & REAGENTS:

  • X-ZELL Cryofixation Station™
  • Cryofixation Buffer I
  • Cryofixation Buffer II
  • Slide fixation cartridges
  • Cryofixation Station

Step 3: Cryoimmunostaining™

Cryoimmunostaining™ is a patented, highly multiplexed immunofluorescence staining method using X-ZELL world-unique Cryoimmunostainer™ (capable of handling a throughput of 60+ slides a day) and a range of proprietary buffers. For perfect results every time, only use X-ZELL’s proprietary, low consumption capillary gap system, CapGap™.

Validated for five visible light, two near-infrared (NIR) and one infrared (IR) channel, Cryoimmunostaining™ has proven to preserve cell morphology unlike any other method currently on the market, to avoid crosstalk and non-specific antibody binding and to re-stain entire slides with a different set of antibodies if required.

Most importantly, it allows rapid imaging of eight-color slides on standard widefield fluorescence microscopes without requiring any further image processing. For visible and near-infrared dyes, analysis can be performed in the eyepiece of the microscope, without any imaging required.

Complete immunostaining workflow (excluding image analysis): 2 hours. Hands-on time: 20 minutes.

 

MATERIALS & REAGENTS:

  • X-ZELL Cryoimmunostainer™
  • Blocking Reagent Set
  • Antibody Binding Buffer
  • Pre-mount Reagent
  • Coverslip Mounting Reagent
  • CapGap™ Capillary Gap slide clips

ADDITIONAL INFORMATION

Stop optimizing, start discovering

MICROSCOPES SUITABLE FOR SLIDE IMAGING:

  • Any widefield or confocal fluorescent microscope equipped with suitable filter sets (contact us for a list of validated fluorophore/filter combinations).
  • Validated microscopes include the Leica DM/DMI Series, the Nikon Te/Ti Series and the Olympus BX and IX series. Widefield fluorescence and confocal microscopes of other manufacturers are suitable as well.
  • Automated microscopes are not required but accelerate slide analysis. Microscopes with less than 8 filter positions can sometimes be upgraded with external filter wheels. Alternatively, in some systems, filter cubes can be changed manually between image acquisition.
  • Monochrome cameras offer higher sensitivity for fluorescence imaging, facilitating very brief exposure times and are therefore preferred over color cameras.

 

To enquire about X-ZELL Inc.’s Cryoimmunostaining™ suite of products, order X-ZELL reagents or CapGap™ equipment, please contact sales@x-zell.com.

To obtain detailed Cryoimmunostaining™ process protocols, please contact info@x-zell.com.

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