 PURCHASE REAGENTS FROM SUPPLIER

 OBTAINS THE REQUESTED REAGENT

 REAGENT HANDLING AND STORAGE
 REAGENT USAGE AND MANAGEMENT
 PERFORMS FLOW CYTOMETRY TESTING
 DIPOSAL AND ELIMINATION OF WASTAGE
 MONITORS MONTHLY INVENTORY
 REQUESITION OF THE REAGENT
 PURCHASING OF THE REAGENT
Purchasing Department personnel handles all
requestion made by the section and facilitates
ordering from the supplier.

DELIVERY OF THE REAGENT

Received by the HPD DELMONTE warehouse staff,
including checking of proper delivery handling and
required transport temperature.
MACHINE OPERATION AND
PROCEDURAL TRAINING
PROCUREMENT OF THE REAGENT CERTIFICATION
Requested on the SAP requisition program and The RMT must be trained and certified
acquired by HEMATOLOGY section staff (RMT/ LAB to perform flow cytometry testing.
PROCESSOR).

REAGENT USAGE AND MANAGEMENT

Used by Hematology RMT for FLOWCYTOMETRY
TESTING (BASIC AND COMPREHENSIVE
LEUKEMIA PANEL)

HANDLING OF THE REAGENT

(1:0) Dilution preparation of the “working” reagent
done by the section RMT prior testing. Freshly
INVERTORY AND REAGENT prepared or stored (excess reagent per batch
REQUISTION running. DIPOSAL AND ELIMINATION OF
Daily monitoring and monthly inventory WASTAGE
done by the RMT

REAGENT STORAGE
• Prepared working reagents stored at 15-25º C
Room temperature in a closed cap-tightly
sealed 50 mL conical tube, stable for 1 month.
• Unopened reagents stored at 2-8º C
Refrigerator temperature, stable until
manufacturer’s expiration date.
A. CYTOPLASMIC I. INCUBATION J. WASHING Q. RUNNING OF
INTRACELLULAR Room temperature and Add 1mL of diluted (1:10) SAMPLES
STAINING keep it in the dark for 10 Phosphate Buffer Solution Read all tubes on the pre-
Addition of CD45 antibodies minutes controlled FACSVIA
to all CT TUBES. machine

B. ADDITION OF H. PERMEABILIZING K. CENTRIFUGATION P. RESUSPENDING OF

SAMPLES SOLUTION AND DECANTATION WASHED SAMPLES
Bone Marrow aspirate or Add 500uL of diluted (1:10) Centrifuge at 200 g for 5 Add 500uL of diluted (1:10)
Whole Blood (Pre-diluted or Permeabilizing reagents mins. Then decant Phosphate Buffer Solution
Un-diluted) supernatant

C. INCUBATION G. CENTRIFUGATION L. PRE-TITRATED O. CENTRIFUGATION

Room temperature and AND DECANTATION ANTIBODY REAGENTS AND DECANTATION
keep it in the dark for 15 Centrifuge at 200 g for 5 Addition of MPO, CD79, Centrifuge at 200 g for 5
minutes mins. Then decant CD3, CD7 antibodies mins. Then decant
supernatant supernatant

D. LYSING SOLUTION F. INCUBATION M. INCUBATION N. WASHING

Add 2mL of diluted (1:10) Room temperature and Room temperature and Add 2mL of diluted (1:10)
FACS Lysing solution keep it in the dark for 10 keep it in the dark for 15 Phosphate Buffer Solution
minutes- CRITICAL minutes
A. SURFACE H. CENTRIFUGATION I. CENTRIFUGATION
INTRACELLULAR AND DECANTATION AND DECANTATION
STAINING
Centrifuge at 200 g for 5 Centrifuge at 200 g for 5
Addition of CD45, ANTI- mins. Then decant mins. Then decant
HLA, CD117, CD33, CD13, supernatant supernatant
CD20, CD10, CD19, CD34
antibodies.

B. ADDITION OF G. WASHING J. RESUSPENDING OF

SAMPLES Add 2mL of diluted (1:10) WASHED SAMPLES
Bone Marrow aspirate or Phosphate Buffer Solution Add 500uL of diluted
Whole Blood (Pre-diluted or (1:10) Phosphate Buffer
Un-diluted) Solution

C. INCUBATION F. CENTRIFUGATION K. RUNNING OF

Room temperature and AND DECANTATION SAMPLES
keep it in the dark for 15 Centrifuge at 200 g for 5 Read all tubes on the pre-
minutes mins. Then decant controlled FACSVIA
supernatant machine

D. LYSING SOLUTION E. INCUBATION

Add 2mL of diluted (1:10) Room temperature and
FACS Lysing solution keep it in the dark for 10
minutes- CRITICAL
PERSON RESPONSIBLE
Patient Care Corporation-AMHSCO
Enterprises BD Biosciences
HPD Purchasing Department personnel
HPD Warehouse staffs
Hematology section’s Registered Medical
technologist with the aide of laboratory
processor
Hematology section’s Registered Medical
technologist
HPD Laboratory Managers
Head of Laboratory Pathologist, Associate
Clinical Pathologist and Flow Cytometry
Analyst Pathologist
Biosafety officer
AMHSCO Enterprises BD Biosciences
application specialist representative
Regulatory Management
DUTIES AND RESPONSIBILITIES
The distributor of BD FACS™ Lysing Solution and provides with reagent inserts, Material Safety Data Sheet,
and Certification of Product Registration.
Handles all the reagent requisitions made by the section and purchases of BD FACS™ Lysing Solution from the
supplier.
Delivery transactions from the supplier to the HPD Hematology section and first-hand reagent receiving
Checking of proper reagent transport temperature, lot number and expiration dates. Newly acquired
reagents will be added to the SAP requisition program.
Manages the requesting (SAP requisition program), monthly inventory, monitoring, storage, and disposal of
the reagent.
Establishment of flow cytometry standard machine operating procedures, work instructions and quick guides
based on reagent inserts and machine operating manual.
Must be trained in flow cytometry testing which includes reagent handling and preparation, test procedural,
flow cytometer machine operation, basic troubleshooting, aliquoting and pipetting training.
Must also attends Biosafety and Risk Assessment training, and proper wearing of PPE (personal protective
equipment) orientation.
Reviews and checking of standard machine operating procedures, quick guides and work instructions.
Approval and validations of standard machine operating procedures, quick guides and work instructions.
Conducts regular Biosafety and Risk Assessment training, aliquoting and pipetting of reagents and samples
training for newly hired RMTs.
Provides and facilitates flow cytometer machine operation and end-user training and certification.
Displays instructional sign boards in the laboratory which includes proper hand washing technique, wearing
of personal protective equipment, biohazard signage and Biosafety Officer’s contact number information.