Module 1-Intro
Module 1-Intro
➢ Special Chemistry - less frequently ordered tests, labor intensive and often manual
methods generally non-stat tests (result not required immediately). Employs
Electrophoresis, HPLC, Infrared Spectroscopy, Radioimmunoassay (RIA), GC-MS
Laboratory Glassware
1. High thermal resistant glass (borosilicate glass with low alkali content – kimax,
pyrex).
2. Alumina-Silicate glass - they are strengthened chemically rather than thermally and
six times stronger than borosilicate example is Corex, they resist clouding and
scratching and alkali resistant.
a. Corex brand – glass which has been strengthened chemically.
b. Vycor brand – recommended for use involving high temperature; drastic heat
shock. It is primarily used in ashing and ignition techniques.
3. High silica glass
4. Glass with higher resistance to alkali
5. Low Actinic Glassware
This contain materials that imparts an amber or red color to the glass and reduce the
amount of light passing to the substance within the glassware's.
It was developed to provide a highly protective laboratory glassware for handling
materials sensitive to light (bilirubin, carotene and Vitamin A).
5. Standard Flint Glass
This is a soda–lime glass composed of mixtures of the silicone, calcium and sodium.
This type of glass is lowest in cost and can readily be fabricated in a variety of shapes.
Glass of choice for weighing bottles.
Glassware can be divided into 2 groups
1. Non-volumetric glassware
Beaker
Flask
2. Volumetric Glassware
Volumetric Flask
Graduated Cylinder
Pipets (Pipettes)
Laboratory instrument used to transport a measured volume of liquid.
Three types of glass pipets used in the laboratory
1. Volumetric - Designed to deliver a single volume precisely, the volume will be
indicated near the top of the pipet, At the top of the pipet is an etched ring.
has a large bulb, and is calibrated for a single volume. Typical volumes are 10, 25, and
50 mL
Fluid must be drawn up the pipet to above the ring indicating the volume and then
released slowly until the bottom of the meniscus is exactly at the ring.
To transfer this volume to a second container, touch the pipette tip to the inside of the
new container and allow the liquid to drain out.
Types of Volumetric pipettes:
a. Ostwald–Folin pipette
A modified volumetric pipette with their bulbs closer to the delivery tip. They are used
for measuring from 0.5 to 10 ml of viscous fluids such as whole blood, serum or plasma.
The pipette has an etched ring near the mouthpiece indicating that the last drop be
blown out (blow–out pipette).
b. Van Slyke pipette
Constructed of thick-walled capillary tubing with a bulb in the center and one mark
above and one below. The specified volume is delivered between these two marks.
2. Graduated Pipet
a. Mohr - TC - sometimes called “drain out pipettes”, a pipet with this marking has been
calibrated to contain a specified volume of liquid. These pipets have a single painted or
frosted ring at the top and are allowed to simply drain with the tip placed against the
side of the receiving vessel.
To accurately transfer fluid with this type of pipet, the meniscus must be precisely on a
calibration mark both at the beginning and at the end of a transfer.
b. Serological - A pipet marked TC, also known as a “blow out” pipet, has been
calibrated "to contain" a specified volume of liquid.
These pipets will have no base mark and graduations continue onto the tip. This means
that ALL the measured liquid in the pipet must be delivered.
A 5 mL serological pipette with one-tenth milliliter graduations can be used to obtain
any desired volume of liquid in one-tenth milliliter increments between 0.1 mL to 5.0mL,
although volumes smaller than 0.2 mL are best measured with automatic pipets.
3. Pasteur Pipettes
Pasteur pipettes are made from glass. With its bulb-shaped top, a Pasteur pipette
resembles an archetypal liquid dropper. Pasteur pipettes are considered fairly
inaccurate today. They are neither calibrated nor graduated, and are more often used in
biology – rather than chemistry – laboratories as a way of transferring aqueous
solutions from one container to another. Named after French physician Louis Pasteur,
Pasteur pipettes are often disposed of after use.
4. Micropipettes – available for measuring very small volumes (0.5 ml or less).
Volumes are often expressed in lambdas. One lambda equals 0.1 ml or 0.001 ul. It is
calibrated either TC or TD. The TC or the wash out pipette are more accurate of the two.
When using micropipettes, the sample is drawn just above the mark, the outer surface
of the pipette is wiped clean with tissue paper and the sample is adjusted to the mark
by touching the tip repeatedly to the paper. The contents are then delivered rinsing
several times into a diluent. The diluent should not be drawn above the calibration mark
because this will add a film of sample in excess of normal value.
a. Sahli hemoglobin pipette – calibrated to contain .02 ml (20ul).
5. Automatic pipettes – dispenses a pre–rated sample when plunger is moved through
its complete stroke. Used to accurately deliver very small volumes, microliters, of liquid.
A volume of 0.1 mL is equal to 100 microliters
a. Peristaltic type b. Piston type c. Seligson type
Beaker
Used for transferring liquid to another container or to transfer a small amount of
reagent for use in procedures. Volume is not accurate, just an estimate.
NEVER PLACE A REAGENT IN ANOTHER CONTAINER WITHOUT LABELING THE
CONTAINER FIRST.
Erlenmeyer Flask
Features a conical base, a cylindrical neck and a flat bottom.
They are marked on the side (graduated) to indicate the approximate volume of their
contents. This is NOT used for ACCURATE measurement
Graduated Cylinder
For rapid measurement of liquid volume. They are generally more accurate and precise
for this purpose than flasks. This is a semi accurate liquid measuring vessel.
Volumetric Flask
A volumetric flask is used to measure very precisely one specific volume of liquid (100
ml, 250 ml, etc., depending on which flask you use).
This flask is used to prepare a solution of known concentration.
To make up a solution, first dissolve the solid material completely, in less fluid than
required to fill the flask to the mark.
After the solid is completely dissolved, very carefully fill the flask to the mL mark.
The top is then sealed and the flask is inverted several times to mix.
Conversion:
Dilutions
A ratio of the concentrate to the total (final) volume.
A 1:4 dilution has a 1 volume of sample and 3 volumes of diluent mixed together.
Any volume can be used to create this dilution, but it must be the same unit of volume
Keep in mind the sample size when making your dilution
For example: a 2:3 dilution could contain:
2 mL serum: 1 mL pure water
20 µL of serum: 10 µL of pure water
0.2 mL of serum: 0.1 mL of pure water
Serial Dilutions
In these types of questions, you are given a series of tubes. Each tube having a
measured amount of a diluent. You are instructed to add a specified amount of
specimen into the first tube, mix well and transfer a specified amount of the mixture to
the next tube, etc.