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.
- 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 INFORMATIONStop 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 email@example.com.
To obtain detailed Cryoimmunostaining™ process protocols, please contact firstname.lastname@example.org.
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