IVD Ready for use FISH Probe Kits Full List

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Our Product Advantages:

a). FISH probes (FastProbe®) hybridization time of 2 hours (blood cell and tissue) against 16-24 hours for conventional probes in the market. 

b). FISH probes (FastProbe®) using the Non-Repetitive Sequences technology ensuing a product with a very good hybridization result, good and clear signal to background noise, high specificity and accuracy.

c). FISH probes (FastProbe®) with a very good anti-quenching effect allowing long observation under fluorescence microscope.

d). An excellent customer service with very short lead time from PO confirmation to shipment.

e). Very competitive affordable price. 

Fast Fluorescence In-Situ Hybridization (FISH) Probe

Protocol Manual

        1. SAMPLE

        A. Tissue

        (1) a). Paraffin-embedded tissue samples fixation with 4% formaldehyde for 6-72 hours.

           b). Thickness: 3-5µm

        (2) Glass slides: Adhesive slides.

         

        B. Cells

        (1) a). Anticoagulated fresh bone marrow (Storage conditions 4^oC, within 24h).

           b). Bone marrow cells accurate fixation (Storage conditions -20^oC, within 6 months).

        (2) Glass slides: Ordinary/Common glass slides.

         

        2. SAMPLE PRETREATMENT 

        Pretreatment procedure

        A. Tissue - Slides pretreatment (sample releasing agent)

        1. Slides heating: Heat at 80^oC for 30min or 65^oC for 2h or overnight.

        2. Dewaxing: Deparaffinization agent 68^oC for 15min.

        3. Washing: Wash with anhydrous EtOH at room temperature for 5min.

        4. Permeation: Permeation agent 90^oC for 20min or deionized water 90^oC for 40min.

        5. Washing: Wash with distilled water at 37^oC for 3min.

        6. Digestion: Enzymic digestion at 37^oC for 10-40min. Enzyme working buffer and Protease solution (10x) in 9:1 proportion.

        7. Washing: At room temperature, wash with cleaning solution twice, 5min each time.

        8. Dehydration: With 70%, 85% and 100% gradient EtOH for 2min.

        9. Dry at room temperature.

         

        B. Cell - Slides pretreatment

        1. Slides heating: Heat at 56^oC for 30min.

        2. Washing: At room temperature, wash with cleaning solution twice, 5min each time.

        3. Dehydration: With 70%, 85% and 100% gradient EtOH for 2min.

        4. Dry at room temperature.

 

        3. DENATURATION & HYBRIDIZATION

        Denaturation

        Before probe use, invert the tube up and down for 5 times. Centrifuge instantaneously for 1-2 sec., if there is a persistent red substance, invert until the substance disappearance.

         

        A. Tissue - Slides FISH hybridization

        a). 85^oC denaturation for 5min.

        b). 42^oC hybridization for 2h (for pretreatment made with HealthCare Biotech FISH Pretreatment Reagent Kit) or Overnight (for pretreatment made without HealthCare Biotech FISH Pretreatment Kit Reagent).

         

        B. Cells - FISH hybridization

        a). 88^oC denaturation for 2min.

        b). 45^oC hybridization for 2h.

         

        4. WASHING

        1. Wash at room temperature with 2xSSC solution for 1min.

        2. Washing solution 68^oC (Neutral 0.3% NP-40/0.4 x SSC solution) for 3min.

        3. Deionized water at 37^oC for 1min.

        4. Dry at room temperature.

         

        5. DYEING

        DAPI counterstaining for 10 min.

         

        6. RESULTS READING

        Target area analysis under fluorescence microscope (HealthCare Biotech FastProbe® filter parameters: Green (495/518), Orange (553/565), DAPI (367/452).