· Pipettes/pipette tips – volumes ranging between 2-1000 uL
· Drying oven (able to reach & hold 60℃)
· Decloaking Chamber NxGen (Biocare Medical DC2012/DC2012-220V)
o ----------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)
o ----------HybEZ oven (ACD 321710/321720)
o ----------Humidity control tray (ACD 310012)
o ----------HybEZ Humidifying Paper (ACD 310025)
o ----------EZ-Batch Wash Tray (ACD 321717)
o ----------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)
o ----------Hydrogen Peroxide (ACD 322335)
o ----------Protease Plus (ACD 322331)
· RNA-Protein Co-Detection Ancillary Kit (ACD 323180)
o ----------Co-Detection Target Retrieval Reagents (ACD 323165/323166)
o ----------Co-Detection Antibody Diluent (ACD 33160)
o ----------Co-Detection Blocker (ACD 323170)
· RNAscope Wash Buffer Reagents (ACD 310091/320058)
· RNAscope Multiplex Fluorescent Detection Reagents v2 (ACD 323110)
o ----------Amp 1 (ACD 323101)
o ----------Amp 2 (ACD 323102)
o ----------Amp 3 (ACD 323103)
o ----------HRP-C1 (ACD 323104)
o ----------HRP-C2 (ACD 323105)
o ----------HRP blocker (ACD 323107)
o ----------DAPI (ACD 323108)
· RNAscope Multiplex TSA Buffer (ACD 322809)
· Opal 690 (Perkin Elmer FP1497A)
· Opal 570 (Perkin Elmer FP1488A)
· Alexa Fluor 488 F(ab’) fragment of goat anti-rabbit IgG (Invitrogen A11070)
· Anti-IBA1 rabbit polyclonal antibody (500ug/mL; Fujifilm Wako 019-19741)
· RNAscope Probe, Channel 1
(interchangeable with other channel 1 probe to detect different transcript)
o ----------CSF1R (ACD 1234601-C1)
· RNAscope Probe, Channel 2 (interchangeable with other channel 2 probe to detect different transcript)
o ----------CD4 (ACD 491891-C2)
· SSC Buffer 20X Concentrate (Sigma S6639-1L)
· ProLong Gold Antifade reagent (Invitrogen P36930)
· #1 thickness cover glass (Fisherbrand 12-545-F)