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Dilutions

Instructor: Cecile Sanders, M.Ed., MT(ASCP), CLS (NCA)

Dilutions for the Clinical Laboratory
Dilution = making weaker solutions from stronger ones Example: Making orange juice from frozen concentrate. You mix one can of frozen orange juice with three (3) cans of water.

in the O. the dilution would be expressed as 1/4. When saying the dilution. you would say.J. to a TOTAL of four cans of diluted O. . for one can of O.Dilutions for the Clinical Laboratory (cont·d) Dilutions are expressed as the volume of the solution being diluted per the total final volume of the dilution In the orange juice example on the previous slide.J.J. example: ´one in fourµ.

. because you express the volume of the solution being diluted (1 ml of serum) per the TOTAL final volume of the dilution (10 ml total). the dilution would be written 1/10 or said ´one in tenµ.Dilutions for the Clinical Laboratory (cont·d) Another example: If you dilute 1 ml of serum with 9 ml of saline.

. The total solution volume is 101 parts (1 part acid + 100 parts water).Dilutions for the Clinical Laboratory (cont·d) Another example: One (1) part of concentrated acid is diluted with 100 parts of water. The dilution is written as 1/101 or said ´one in one hundred and oneµ.

Dilutions for the Clinical Laboratory (cont·d) Notice that dilutions do NOT have units (cans. ml. or parts) but are expressed as one number to another number Example: 1/10 or ´one in tenµ .

If more than one part of original substance is initially used.Dilutions for the Clinical Laboratory (cont·d) Dilutions are always expressed with the original substance diluted as one (1). . it is necessary to convert the original substance part to one (1) when the dilution is expressed.

use a ratio and proportion equation. .0___ ______2 parts dye 10 parts total volume x 2x = 10 x = 5 The dilution is expressed as 1/5. The total solution volume is 10 parts (2 parts dye + 8 parts diluent). but the original substance must be expressed as one (1). remembering that dilutions are stated in terms of 1 to something: = ___1. The dilution is initially expressed as 2/10.Dilutions for the Clinical Laboratory (cont·d) Example: Two (2) parts of dye are diluted with eight (8) parts of diluent (the term often used for the diluting solution). To get the original volume to one (1).

Example: Two parts (2) parts of whole blood are diluted with five (5) parts of saline.5 . remembering that dilutions are stated in terms of 1 to something: = ___1.Dilutions for the Clinical Laboratory (cont·d) The dilution does not always end up in whole numbers. this is calculated by using the ratio and proportion equation. The dilution would be 2/7. more correctly.0___ __2 parts blood_____ 7 parts total volume x 2x = 7 x = 3. The total solution volume is seven (7) parts (2 parts of whole blood + 5 parts saline). Again. or. 1/3.5 The dilution is expressed as 1/3.5.

The result (answer) using the diluted sample must be multiplied by the RECIPROCAL of the dilution made. . The RECIPROCAL of a 1/5 dilution is 5.Dilutions for the Clinical Laboratory (cont·d) Dilution Factor ² used to correct for having used a diluted sample in a lab test rather than the undiluted sample.

it is necessary to multiply the result by the dilution factor (in this case x 2). The final result is 210 g/dl x 2 = 420 g/dl. .Dilutions for the Clinical Laboratory (cont·d) Correction for using a diluted sample Example: A technician performed a laboratory analysis of patient·s serum for a serum glucose (blood sugar) determination. To correct for the dilution. The patient·s serum glucose was too high to read on the glucose instrument. The technician diluted the patient·s serum 1/2 and reran the diluted specimen. obtaining a result of 210 g/dl.

A dilution of the stronger solution of substrate is needed.Dilutions for the Clinical Laboratory (cont·d) Sometimes it is necessary to make a dilution of an existing solution to make it weaker. . Example: A 100 mg/dl solution of substrate is needed for a laboratory procedure. All that is available is a 500 mg/dl solution of substrate.

Dilutions for the Clinical Laboratory (cont·d) To make a weaker solution from a stronger one. use this formula: V1 x C1 = V2 x C2 Example: To make 100 ml of the 100 mg/dl solution from the 500 mg/dl solution needed in the previous example: V1 = 100 ml V2 = V2 (unknown) C1 = 100 mg/dl C2 = 500 mg/dl 100 ml x 100 mg/dl = V2 x 500 mg/dl V2 = 20 ml Dilute 20 ml of 500 mg/dl solution up to 100 ml with water to obtain 100 ml of 100 mg/dl substrate solution .

A general rule for calculating the dilution of solutions obtained by diluting in a series is to MULTIPLY the original dilution by subsequent dilutions.Serial Dilutions Dilutions can be made singly (as shown previously) or in series. . in which case the original dilution is diluted further.

Serial Dilutions (cont·d) Example of a serial dilution: .

Serial Dilutions (cont·d) In the serial dilution on the previous slide. . 1 ml of stock solution is mixed with 9 ml of diluent. but the concentration of stock in the second tube is 1/10 x 1/10 for a 1/100 dilution. for a 1/10 dilution. Then 1 ml of the 1/10 dilution is mixed with another 9 ml of diluent. The second tube also has a 1/10 dilution.

in the third tube. Again you have a 1/10 dilution in the third tube. but the concentration of stock in the third tube is 1/10 x 1/10 x 1/10 for a 1/1000 dilution.Serial Dilutions (cont·d) Continuing with the serial dilution. This dilution could be carried out over many subsequent tubes. you mix 1 ml of the 1/100 dilution from the second tube with 9 ml of diluent in the third tube. .

where technicians need to make dilutions of patient·s serum to determine the weakest concentration that still exhibits a reaction of some type. The RECIPROCAL of the weakest concentration exhibiting a reaction is called a ´titerµ. .Serial Dilutions (cont·d) Serial dilutions are most often used in serological procedures.

but NOT in tubes 4 or 5. The titer = 1000.000 Tube #5 = 1/100. .000 Reactions occur in tubes 1 through 3.Serial Dilutions (cont·d) Example of determining a titer: A technician makes a serial dilution using patient serum: Tube #1 = 1/10 Tube #2 = 1/100 Tube #3 = 1/1000 Tube #4 = 1/10.

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