SCYM ASCP UPDATED TEST PAPER QUESTIONS
AND ANSWERS SURE A+
✔✔Sample Identification and Handling - ✔✔A biosafety assessment of some type must
be carried out by the facility to assure that the operator will have specific knowledge of
the origin of the sample and awareness of the potential presence
of pathogens or genetically modified material. This is critical to protection of the
operator. A bio safety information sheet or questionnaire on the specimens will be
completed and approved
by the flow cytometry laboratory manager prior to the start of any new experiments. An
example is provided as an appendix to this document, for implementation at each
individual core or
laboratory.
The appropriate biosafety containment level must be applied in relation to the nature
and status (e.g. live vs. fixed cells) of the sample.
Guidelines on handling and proper disposal of biological materials must be followed in
all flow cytometry laboratories.
✔✔Personal Protective Equipment - ✔✔Operators must wear appropriate PPE for the
procedures and the materials to be analyzed or sorted. This includes at a minimum,
gloves, lab coat or disposable Tyvek gown and eye protection.
Work with certain materials may also require respiratory protection (in the form of N-95,
N-99 or N-100 respirators or PAPRs) in addition to the use of biological safety cabinets
for containment.
✔✔BioSafety Requirements - ✔✔For the purposes of working with any human materials
and certain animal materials (to include tissues, primary and immortalized cell lines)
those materials are defined as Risk Group 2 (RG2)
materials for laboratory manipulations, which must be conducted with Biosafety Level 2
containment and practices.
BSL 2 containment also consists of the work practices (good microbiological laboratory
technique) and when appropriate, a certified, properly operating biosafety cabinet
, (BSC), typically Class II, Type A, or Type A/B3. Work will be performed in a biosafety
cabinet (BSC) as appropriate.
✔✔Laser Hazzard - ✔✔Lasers are used in many of these instruments. With few
exceptions, these machines include hard to-disable safety devices that prevent user
exposure to the beams in normal use. Analytical flow cytometers only present a laser
hazard if the safety covers are removed or the interlocks are defeated. Users are
reminded not to stare at the objective/specimen while the instrument is scanning.
✔✔Sorting Unfixed Cells - ✔✔All manipulations of unfixed human specimens must be
conducted in accordance with the University policy on Bloodborne Pathogens/Exposure
Control. Unfixed biological specimens of
human or animal origin can harbor known and unknown pathogens that may be
transmitted through droplets and aerosols that are generated during the cell sorting
process. This requires handling all unfixed human specimens as potentially infected
with HIV, Hepatitis B and/or Hepatitis C. This also requires the PI to assure personnel
are trained in the required procedures with strict adherence to the Standard Precautions
described in the University policy.
SOP must be made for the handling of the following:
-Unfixed human materials from patients (blood samples, various body fluids, cultured
cells and environmental samples) shall be considered as potentially infectious, per the
OSHA
Bloodborne Pathogens Standard and University policy (e.g. peripheral blood leukocytes,
bone marrow, etc.)
-Unfixed cells from primary or immortalized cultures from human donors or of human
origins (e.g. HELA cells, tumor cells lines, etc)
-Unfixed cells from primary and immortalized cultures from animal donors, to include
nonhuman, primates, other species, and transgenic animals.
-CD34 stem cells, fused dendritic cells + tumor cells, tumor cells which have been
transfected with rDNA or synthetic nucleic acid molecules, It is incumbent upon the
laboratory (Principal Investigator or other research
✔✔Flow Cytometry BioHazzards - ✔✔If a sample contains lentivirus, adenovirus, or any
other genetically engineered amphotropic virus, it must be treated as a potentially
infectious hazard and all Biosafety Level 2 practices and
procedures must be followed. This means that any specimens must be kept closed at all
times except when being put onto the
cytometer sample introduction port.
Vortexing of open tubes is always prohibited. Vortexing of any closed tube should be
done in a biological safety cabinet, if available. Otherwise, personnel must take great
care when vortexing
and when opening vortexed materials.
High speed sorting generates aerosols. Typically, only fixed materials may be sorted in
a cellsorter, not otherwise contained in a biological safety cabinet.
AND ANSWERS SURE A+
✔✔Sample Identification and Handling - ✔✔A biosafety assessment of some type must
be carried out by the facility to assure that the operator will have specific knowledge of
the origin of the sample and awareness of the potential presence
of pathogens or genetically modified material. This is critical to protection of the
operator. A bio safety information sheet or questionnaire on the specimens will be
completed and approved
by the flow cytometry laboratory manager prior to the start of any new experiments. An
example is provided as an appendix to this document, for implementation at each
individual core or
laboratory.
The appropriate biosafety containment level must be applied in relation to the nature
and status (e.g. live vs. fixed cells) of the sample.
Guidelines on handling and proper disposal of biological materials must be followed in
all flow cytometry laboratories.
✔✔Personal Protective Equipment - ✔✔Operators must wear appropriate PPE for the
procedures and the materials to be analyzed or sorted. This includes at a minimum,
gloves, lab coat or disposable Tyvek gown and eye protection.
Work with certain materials may also require respiratory protection (in the form of N-95,
N-99 or N-100 respirators or PAPRs) in addition to the use of biological safety cabinets
for containment.
✔✔BioSafety Requirements - ✔✔For the purposes of working with any human materials
and certain animal materials (to include tissues, primary and immortalized cell lines)
those materials are defined as Risk Group 2 (RG2)
materials for laboratory manipulations, which must be conducted with Biosafety Level 2
containment and practices.
BSL 2 containment also consists of the work practices (good microbiological laboratory
technique) and when appropriate, a certified, properly operating biosafety cabinet
, (BSC), typically Class II, Type A, or Type A/B3. Work will be performed in a biosafety
cabinet (BSC) as appropriate.
✔✔Laser Hazzard - ✔✔Lasers are used in many of these instruments. With few
exceptions, these machines include hard to-disable safety devices that prevent user
exposure to the beams in normal use. Analytical flow cytometers only present a laser
hazard if the safety covers are removed or the interlocks are defeated. Users are
reminded not to stare at the objective/specimen while the instrument is scanning.
✔✔Sorting Unfixed Cells - ✔✔All manipulations of unfixed human specimens must be
conducted in accordance with the University policy on Bloodborne Pathogens/Exposure
Control. Unfixed biological specimens of
human or animal origin can harbor known and unknown pathogens that may be
transmitted through droplets and aerosols that are generated during the cell sorting
process. This requires handling all unfixed human specimens as potentially infected
with HIV, Hepatitis B and/or Hepatitis C. This also requires the PI to assure personnel
are trained in the required procedures with strict adherence to the Standard Precautions
described in the University policy.
SOP must be made for the handling of the following:
-Unfixed human materials from patients (blood samples, various body fluids, cultured
cells and environmental samples) shall be considered as potentially infectious, per the
OSHA
Bloodborne Pathogens Standard and University policy (e.g. peripheral blood leukocytes,
bone marrow, etc.)
-Unfixed cells from primary or immortalized cultures from human donors or of human
origins (e.g. HELA cells, tumor cells lines, etc)
-Unfixed cells from primary and immortalized cultures from animal donors, to include
nonhuman, primates, other species, and transgenic animals.
-CD34 stem cells, fused dendritic cells + tumor cells, tumor cells which have been
transfected with rDNA or synthetic nucleic acid molecules, It is incumbent upon the
laboratory (Principal Investigator or other research
✔✔Flow Cytometry BioHazzards - ✔✔If a sample contains lentivirus, adenovirus, or any
other genetically engineered amphotropic virus, it must be treated as a potentially
infectious hazard and all Biosafety Level 2 practices and
procedures must be followed. This means that any specimens must be kept closed at all
times except when being put onto the
cytometer sample introduction port.
Vortexing of open tubes is always prohibited. Vortexing of any closed tube should be
done in a biological safety cabinet, if available. Otherwise, personnel must take great
care when vortexing
and when opening vortexed materials.
High speed sorting generates aerosols. Typically, only fixed materials may be sorted in
a cellsorter, not otherwise contained in a biological safety cabinet.
