week one HELP

wed 9-11

immunology and molecular

didactic and student lab

75 or higher

sim lab and clinical practicum

85 or higher

academic integrity

notify student within 10 bus days and referred to PCC, 0%, reported to OIA and investigated, could lose spot in program

approved absent

request 72 hrs in advanced

ASCP

certifies lab pros and continues education, run by pathologists and lab pros, BOC and 3 year maitainence

ASCLS

supports lab mission, run by non physician lab pros, code of ethics and pledge to pro

NAACLS

certifies lab educational programs

certification

non gov just educational requirements passing boards

liscense

state given authority to practice

OSHA mandated safety measures

chemical hygene plan CHP, hazard communication standard HCS, Global harmonizing standard GHS, Bloodborne pathogen plan BBP, exposure control plan ECP, PPE

OSHA CHP

policies to protect employee for handle use and store, update annually, chem inventory, labeling/record keeping, waste disposal, training, PPE, SDS (16)

OSHA GHA

name, pictogram, manufacturer info, signal word, precautionary statement, hazard (severity) statement 1-4 (1 most hazardous) ABC A most severe

HMIS

color bars, health blue, flammability red, reactivity yellow, PPE white, 0-4 0 is least hazardous

NFPA

color diamond, red-yellow same as HMIS, white- specific or special, 0 is least

OSHA BBP

exposure control- determine risk of exposure, standard precations- treat all as infectious, hazard comm standard, Hep B vaccine within 10 days,

pathogens in blood can cause

HIV and HBV

exposure controls

elimination, substitution, engineering (provided by facility to reduce risk), administrative (steps you take), PPE

bio safety cabinet BSC

protects from bioaerosols in micro plating using HEPA filtered laminar air flow, class I II III, UV for disinfection, SASH window

disinfection

kill or reduce pathogenic microorganisms, not spores, 1 part bleach 9 part tap water, before after shift, spill.

disinfection after spill

choose proper PPE according to SDS, contain with paper towels, wet with 10% bleach working from outside in, sit 10-60 min, sweep, kevlar gloves for glass, dispose of waste,

transporting specimen

primary container tube outside is contaminated always touch with gloves, 2ndary container bag outside is uncontaminated, pneumatic tube system- specimen in transport bags foal lined carriers to avoid leaks and hemolysis do not send tissue or biopsy

transport bag

mark with biohazard symbol, 1 specimen type per bag, 1 patient per bag

cap removal and centrifuge

done with gauze and left in gauze

tupes capped, allow to stop on its own, if tube breaks leave lid closed for 30 min

donning and doffing

n- wash hands, gather PPE, gown, eye, gloves, bag, tablet

f- remove tablet, gloves, gown, wash hands, face, wash hands

biosafety levels

bsl- 1-4, 1 least hazardous, 2 most common, bsc III glove box

most common LAIs

not following rules, spills, needles

transmission precautions

contact- gloves and gown, droplets- mask, droplets- N95 resp, neg pressure room

labeling containers

name, precaut and hazard info, date mixed, concentration, expiration

storing acids and bases

away from each other

a- organic away from inorganic, oxidizing away from organic, flammable, and reducing agents

b- inorganic hydroxides in polyethylene

brightfield microscope

light passes from a source through a condenser and is magnified by 2 magnifying lenses the objective and the eyepiece (ocular) together = total magnification

framework

base arm stage

illumination system

light source, condenser (focus light onto specimen), aperture iris diaphragm (controls angle and amount of light), field iris diaphram

focusing system

interpupillary distance adjustment (space between eyes), left ocular (diopter) adjustment- focus uneven vision, body tube motion (course- moves stage only use with 10x and fine focus with 40 and 100)

objectives

10x- low power dry field

40x- high power dry

100x- oil

wet prep slide vs stain fixed

10x- low light intensity 40x medium, slightly open diaphragm

10x- low 40x- medium 100x- high, fully open

Birefringence

object that rotates or polarizes light

Numerical aperture (NA)

a mathematically derived number that indicates the resolving power of a lens system

resolution

the ability of a lens to distinguish two points or objects as being separate

parfocal

objectives that remain in focus when switching from one objective to another of differing magnification.

sf in 600

1

sf in 22.840

5

kilo

1000 or 10 3

deci

0.1 or 10 -1

milli

0.001 or 10-3

micro u

0.000001 or 10-6

nano

0.000000001 or 10-9

K to C

F to C

C+ 273

5/9 (F-32)

concentration

amount of solute in given solution

ratio/proportions

concentrations of starting and ending solution are the same

if starting(stock) and ending(work) solution concentrations change

C1V1=C2V2

solute vs solvent(diluent)

mass vs volume (total volume-solute)

normality

# of eq/L, valance- # OHs or Hs, Molarity x valance

dilution

weakend solution, solution (concentrated solute), + diluent (solvent)

1:4

3 parts diluent

serial dilution

use new dilution as stock for a following dilution, fold- tube dilution are the same, solution(final) is diff for each tube

solution dilution

n= factor number (n-fold), x= tube #

titer

most diluted tube with a positive result

multiple (discrete) dilutions

from og sample, not same dilu

class A vs B equipment according to ASTM and ISO

a- ASTM garantee, most precise and accurate, individually calibrated, Lower error tolerance, used for analytical/quantitative testing

b-batch calibrated, used for routine tasks, higher error tolerance, cheaper, durable

TC

The vessel contains the specified volume, but may not deliver it accurately, you must get every last drop out

TD

The vessel is calibrated to deliver the specified volume when allowed to

drain freely, residual liquid left in the tip is intentional and accounted for

serologic pipettes

Long tubes, graduations extending to the tip, measure and deliver, TD, Ideal for reagents or diluents in volumes 1-25 mL, “Blow out” pipettes- frosted rings or double strips, safety bulb expels the final drop

volumetric pipettes

Open-ended bulb in center holds bulk of fluid, more accurate than serological, measure and transfer specific single volumes (preparing calibrators and standards), TD, no volume graduations to the tip, sizes 5 and 10 mL most common, Self-draining- blowing out resid will over transfer, greatest accuracy

micropipettes

Single-hand use thumb goes on plunger Automatic, Fixed volume- delivers one specific volume, repetitive limited testing, Variable volume- indicator dial to adjust delivery, more versatile but higher cost, Single-stop- depress plunger 1x all in one motion, Double-stop- depress plunger 2x, 2 steps to dispense liquid

small volumes ranging from 1- 5000 μL, multiplies the inherent error

forward vs reverse pipetting

for aqueous solutions like serum, plasma, and urine

for viscous, dense, or volatile solutions (e.g., whole blood, alcohols) to prevent delivery errors

pre-wetting

aspirate and dispense the liquid once or twice before the actual measurement to improve accuracy and pre-wet the tips

barrier pipettes and tips

filter that acts as a liquid and aerosol barrier, prevents cross-contamination between samples and protects the internal equipment, Essential for sensitive techniques like Polymerase Chain Reaction (PCR) to prevent DNA amplification of contaminants

stability

Time window for test to be accurately performed

labile

Analytes or organisms that are time or environment sensitive

temp C

room- 20-25, fridge 2-8, freezer -10, incubator 35-37

Aliquot

A portion of a specimen transferred into one or more aliquot/transfer tubes, labeling is critical

labware comp func and accuracy

c- plastic or glass

f- containers and receivers or volumetric TD or TC

a- class a or b

comp- plasticware

cheaper, durable, good for alkaline solutions, leaching into solutions, permeability to water vapor, evaporation, absorbs dyes or proteins

comp- glassware

higher chem stability, clearer, good for analytical work, leach/permeable with some agents

types- borosilicate (most common), alumina-silicate, acid resistant, alkali resistant, low actinic, flint, disposable

func- containers and receivers

beakers, erlenmeyer flasks, test tubes, and reagent bottles

func- volumetric

flasks, graduated cylinders, pipettes

balances precision/top-loading vs analytical

p- measure heavy samples, less accurate, general measurements and bulk weighing

a- measure lighter/minute samples, highly accurate(glass shield protects against air and dust), used for chemical analysis and pipette calibrations

centrifuges

separates particles in a mixture by size and density, precipitate- sediment packed at bottom of tube, supernatant- liquid at top of tube, process blood and prepare urinary sediment

water for purification

distillation- remove minerals, metals, bacteria using heat to vapor and condense, deionization (ions through ion exchange calcium sodium chloride), reverse osmosis (force water through semipermeable membrane, metals, minerals, dissolved solids)

water ASTM / old CLSI

I- most pure, standard solutions bufferes and controls, quantitative analytical procedures, electrophoresis, toxicology screening tests, and liquid chromatography

II- qualitative chem and hemato immun micro

III- qualitative urinalysis, washing labware

IV- least pure

water new CLSI

CLRW- replace I and II, used for patient testing and preping reagents

SRW- ultra pure, used for specialized testing- molecular

IFW- specification by manufacturers, for automated lab analyzer

saline

salt (NaCl) and water, physiologic- .85-.9% concentration dilute specimen, buffered- maintains pH and osmotic pressure (PBS or FMH), sterile- free of microorganisms for identifying tests in micro

reagent labeling

name, lot number, expiration date, storage info, strength of solution, hazard statement, preparers name and date

ACS

gold standard reagent

reagent

basically as good as ACS

USP

not as good as ACS but close

NF

not as good as USP but pretty good

laboratory reagent

good not great tho

purified reagent

meh

tech reagent

bad

kit reagents

for immunoreagents or enzymatic reagents, maintain quality control, do not use reagents from diff lot numbers of kits together

blood

wbc, rbc, platelets, plasma

ven- vein, cap- skin, central line or PICC- peripherally not cath, art-art

urine

from kidneys in bladder expel trhough urethra

midstream clean- skin cleansed before collection by patient, cath- tube collects from bladder, supra- Aspiration via needle from bladder, void- skin not cleansed before by patient

fluid specimen

Pleural- lungs, Synovial- joints, Pericar- heart, Peritoneal- abdom organs, Amniotic- fetus

other specimen

Bronchoalveolar lavage lung (BAL)- fluid squirted into lung, sputum lungs- mixture of saliva and mucus, Nasopharyngeal- Upper throat (pharynx) behind nose, Stool/feces, Genital, Abscess- pus in tissue, Tissue- biopsy, Wound- superficial / internal tissue, Saliva

specimen collection

Blood Tubes- Whole blood, plasma, or serum

Specimen Containers- Leak-proof/screw cap, Non-blood containers, Sterile/non-sterile

Devices- Swabs, syringes, transport media

preanalytical phase

test ordered, collection (paitent name, DOB or MRN, date and time, collector name BB must also have SSN and preprinted identifier), transport (STAT, ASAP, Timed, Fasting, Routine), process (req and label match), accept or reject

clotted blood

red- activator promotes clotting, cent- serum (clot 30 min before centrifuge)

gold SST- clot activator and gel, serum separator tube, cent- serum (30 min)

whole blood

green- heparin anticoagulant preventing clotting, cent- plasma (fibrinogen)

green PST- heparin anticoagulant and gel, plasma sep tube, cent- plasma (fib)

blue- sodium citrate anticoagulant prevents clot, cent- plasma (fib)

lavender- EDTA anticoagulant prevent clot, no cent but plasma if done