· Pipettes/pipette tips – volumes ranging between 2-1000 uL
· Drying oven (able to reach & hold 60℃)
· Decloaking Chamber NxGen (Biocare Medical DC2012/DC2012-220V)
----------Can substitute with hot plate by using alternative target retrievalprotocol; see Appendix B: Manual Target Retrieval from AdvancedCell Diagnostics [ACD] FFPE Sample Preparation and Pretreatment User Manual (DocumentNo. 322452)
· Slide staining tray (e.g. Simport M920-2)
· HybEZ II Hybridization System with ACD EZ-Batch Slide System (ACD 321710/321720)
----------HybEZ oven (ACD 321710/321720)
----------Humidity control tray (ACD 310012)
----------HybEZ Humidifying Paper (ACD 310025)
----------EZ-Batch Wash Tray (ACD 321717)
----------EZ-Batch Slide Holder (ACD 321716)
· Tissue-Tek Vertical 24 slide rack (American Master Tech Scientific LWS2124)
· Tissue-Tek Staining Dishes (American Master Tech Scientific LWS20WH)
· Tissue-Tek Clearing Agent Dishes, xylene resistant (American Master Tech Scientific LWS20GR)
***For all reagents, refer to MSDS to determine appropriate precautions, personal protective equipment (PPE), and disposal methods before use***
· Distilled water (obtained in-house)
· 0.05% PBS-Tween (PBS-T), pH 7.35 (made in-house)
· Xylenes (Macron Fine Chemicals 8668-16)
· 100% ethanol (Pharmco 111000200)
· 10% NBF (3.7% formaldehyde; Cancer Diagnostics, Inc. 111)
· ImmEdge Hydrophobic Barrier Pen (Vector H-4000)
· RNAscope H2O2 & Protease Plus Reagents (ACD 322330)
----------Hydrogen Peroxide (ACD 322335)
----------Protease Plus (ACD 322331)
· RNA-Protein Co-Detection Ancillary Kit (ACD 323180)
----------Co-Detection Target Retrieval Reagents (ACD 323165/323166)
----------Co-Detection Antibody Diluent (ACD 33160)
----------Co-Detection Blocker (ACD 323170)
· RNAscope Wash Buffer Reagents (ACD 310091/320058)
· RNAscope Multiplex Fluorescent Detection Reagents v2 (ACD 323110)
----------Amp 1 (ACD 323101)
----------Amp 2 (ACD 323102)
----------Amp 3 (ACD 323103)
----------HRP-C1 (ACD 323104)
----------HRP blocker (ACD 323107)
----------DAPI (ACD 323108)
· RNAscope Multiplex TSA Buffer (ACD 322809)
· Opal 570 (Perkin Elmer FP1488A)
· Alexa Fluor 488 F(ab’) fragment of goat anti-mouse IgG (Invitrogen A11017)
· Anti-Ki-67 antibody; mouse IgG1; clone B56; stock concentration 250 ug/mL (BD 550609)
· RNAscope Probes, Channel 1 (interchangeable with other channel 1 probes if desiring to target other transcripts)
----------MKI67 (ACD 470841)
· ProLong Gold Antifade reagent (Invitrogen P36930)
· #1 thickness cover glass (Fisherbrand 12-545-F)