Immunologic testing is done in many areas of the clinical laboratorymicrobiology, chemistry, toxicology, immunology, hematology, surgical pathology, cytopathology, immunohematology blood bankingand a great variety of specimens are tested. Worlds best powerpoint templates crystalgraphics offers more powerpoint templates than anyone else in the world, with over 4 million to choose from. Good practice for those who have never used a syringe too. If dilution is necessary, apparatus such as volumetric pipettes or airdisplacement pipettes gilson pipettes or equivalent should be used to perform the dilution. A log dilution is a tenfold dilution, meaning the concentration is decreased by a multiple of ten. Discard the pipette in the biowaste disposal containers at the front of. Dilution and concentration theory dilution and concentration theory sounds ominous but you use dilutions and concentrations fairly frequently outside of your scientific life. To complete a tenfold dilution, the ratio must be 1. The experimental method detection limit mdl for tcp was 0. It is used for serological diagnosis of syphilis and it is an example of slide flocculation test. Serial dilutions contd in the serial dilution on the previous slide, 1 ml of stock solution is mixed with 9 ml of diluent, for a 110 dilution. In an experiment involving concentration curves, you can use a serial. Set up the problem dilution factor final volumealiquot volume 0.
Principle, procedure, result interpretation and application principle. Pdf estimation method for serial dilution experiments. The initial concentration and target range needed determines the size and number of dilution steps required. Abo antibody titration madison area technical college. Solution preparation 14 a serial dilution is simply a series of simple dilutions which amplifies the dilution factor quickly beginning with a small initial quantity of material. Tube 2 will be a 12 dilution, 4 will be a 14 dilution. This procedure can be used to establish a standard curve from a solution of known concentration. The pourplate technique is the procedure usually employed. You will make several dilutions of a bacterial stock culture. Ppt dilutions powerpoint presentation free to download. Vdrl stands for venereal disease research laboratory test. As shown in the illustration below, it begins when 1 ml of the bacterial sample is added to 9 ml, and it is mixed together creating a 101 dilution. Using isotonic saline prepare serial dilutions of the test sample positive in the qualitative method 1.
A serial dilution is any dilution in which the concentration decreases by the same factor in each successive step in serial dilutions, you multiply the dilution factors for each step. We worked through the first example together on the board and the students did the dilution using coloured water. This video demonstrates how to quantitate bacteria using the serial dilution and plate count method. Serial dilution method protocol step wise explanation youtube. A small amount of serum or solute can be serially diluted by transferring aliquots to diluent. Samples are serially diluted to the point of extinction, that is, to a point where there are no more viable microorganisms. For example, a tenfold serial dilution could result in the following concentrations.
The titre is reported as the reciprocal of the highest dilution, which shows a positive test result. Serial dilutions austin community college district. For example, dilution factor 10 if a standard curve includes values between 0. The source of dilution material for each step comes from the diluted material of the previous. This dilute solution has 1ml of extract 10ml, producing a 10fold dilution. Label six nutrient agar plates the same as the tubes. Continue doing this from tube to tube as shown in fig. Announcement next test test 2 is on march 22nd, 2011 tuesday at the big hall 8pm test 4 is on april 5th, 2011 tuesday at the big hall 8pm.
To accomplish this, the serial dilutionagar plate technique is used. Analyze the neat samples and the serial dilutions in duplicates, in the same run, and compensate for the dilution factor. The 10 represents the total size of the final sample. Thus, a series of 5, one to ten dilutions equals a one to one hundred thousand dilution. Serial dilutions tube method principle serial dilution is a common technique used in many immunologic procedures. Basic serologic laboratory techniques clinical gate. A small amount of serum or solute can be serially diluted by transferring aliquots to. Announcement next test test 2 is on march 22nd, 2011 tuesday at the big hall 8pm test 4 is on april 5th, 2011 tuesday at the big hall 8pm 2. Serial dilutions serial dilutions are often performed when titering antibodies or when generating amplified dilutions of an analyte. The serial dilution method of bacteria enumeration 1 the serial dilution method of bacteria enumeration 2. Serial dilution pay attention as im going to quiz you on this today 3. Serial dilution, also called limiting dilution series, is a standard laboratory procedure employed to collect the appropriate data in order to accomplish this estimation. Serial dilution is the property of its rightful owner.
Choose a volume for the working stock that you can pipette accurately. Serial dilutions involve diluting a stock or standard solution multiple times in a row. Then 1 ml of the 110 dilution is mixed with another 9 ml of diluent. Each microfuge will intially contain 900 microliters of ppt media in which each microfuge will recieve 100 microliters of the previous solution. When youre thinking about dilution, it helps to simplify your actions into dilution factors. This general microbiology practical lecture explains the serial dilution techniques in pour plate method to isolate bacteria. Label 2 or 3 refer to the firms outline of production op plates per dilution series for each serial. A serial dilution is a stepwise and geometric series of dilutions which starts with a small amount of starting material and amplifies the dilution factor serially by using diluted material as a. A serial dilution is the dilution of a sample, in 10fold dilutions.
In this tutorial, i have explained how to make an animated serial dilution slide in powerpoint. Definition and procedure explained 2012serialdilutionprotocolpdf. When we said the diluted coffee was 110th as strong as the original that was a dilution factor. To detect the end point, multiple serial dilutions are inoculated into a suitable growth medium, and the development of some recognizable. This particular video shows the basics behind setting up and calculating a serial dilution. Read the titre in the last dilution step with visible agglutination and the approximate concentration of the rheumatoid factor can be determined as follows. Once diluted, the suspensions are placed on suitable nutrient media. Working with billions of tiny cells can pose a problem when you need to count the total number of cells in a sample. Serial dilution is a common technique used in many immunologic procedures. For example, a sample size of 1 ml is added to 9 ml of diluent to equal a total of 10 ml. Follow serial dilution technique inoculate labeled empty petri dish with specified ml 0.
Ppt serial dilution powerpoint presentation free to. Bacteria are serially diluted and plated onto petri plates using aseptic technique. In a serial dilution, the final total dilution is a product of each individual dilution in the series. The aoac usedilution methods do not provide procedures to enumerate the test microbe on. Label one tube control and label the remaining five tubes consecutively from 101 through 105. Statistical design for a small serial dilution series. As stated earlier, this method is commonly used to obtain pure cultures of those microorganisms that have not yet been successfully cultivated on solid media and grow only in liquid media. Sop for preparation of culture dilutions for analysis. Standard operating policyprocedure page 5 of 16 standard bacterial plate count uncontrolled copy 3. Theyll give your presentations a professional, memorable appearance the kind of sophisticated look that todays audiences expect. Use the same pipette to add 9 ml of distilled water to a second 15 ml culture tube. Extend this procedure to perform longer serial dilutions. Serial dilution instructions api biotechnology solutions.
This is called a dilution series or a serial dilution. The two most widely used methods for determining bacterial numbers are. This process may be repeated as many times as necessary to achieve the desired solution. For example, when you are making orange juice from a frozen concentrate, you mix 1 can of concentrate with 3 cans of water. Winner of the standing ovation award for best powerpoint templates from presentations magazine. A microorganism that predominates in a mixed culture can be isolated in pure form by a series of dilutions. Use a clean, new, serological transfer pipette to add 10 ml of distilled water to a empty, clean, new, 15 ml culture tube with a cap. It is important to understand the concept of dilutions, since they are a handy tool used throughout all areas of the clinical laboratory. The serial dilution method of bacteria enumeration the serial dilution method of bacteria enumeration many studies require the quantitative determination of bacterial populations.
The serial dilution method of bacteria enumeration ppt video. Notes on titering antibodies lerner research institute. They are described as ratios of the initial and final concentrations. Serial dilutions tube method principle serial dilution is a. Serologic testing has long been an important part of diagnostic tests in the clinical laboratory for viral and bacterial diseases. Typically, the dilution factor remains constant for each dilution, resulting in an exponential decrease in concentration. Powerpoint tutorial no 316 animated serial dilution method in. Perform the qualitative test procedure using each dilution as test specimen. The second tube also has a 110 dilution, but the concentration of stock in the second tube is 110 x 110 for a 1100 dilution. Serial dilution protocol pdf it is a method of diluting a stock solution where concentration decreases by the same quantity in each successive step. Ppt the serial dilution method of bacteria enumeration.
Serial dilutions mlab 35 immunologyserology terry kotrla introduction many of the laboratory procedures involve the use of dilutions. Many studies require the quantitative determination of bacterial populations. A simple method is presented to estimate the microbial counts obtained with the serial dilution technique for microorganisms that can grow on bacteriological media and develop into a colony. A twofold dilution reduces the concentration of a solution by a factor of two that is reduces the original concentration by one half. Pour plate technique procedure, advantages, limitations. Store control cards or serum samples according to the manufacturers directions 3 0.
We could also have said the dilution factor was 110, or the dilution factor was 0. Our designs for this experiment must maximize the probability that useful data will be obtained since these experiments are costly, labor intensive, and time consuming. Learn serial dilutions with free interactive flashcards. Serial dilution ppt 2019 free download as powerpoint presentation. Pipette 90ul of ppt media in each tube x4 pipette 10ul of tetrahymena stock culture into the first tube 101 use the vortex mixer to mix the tetrahymena culture and ppt media thoroughly. Serial dilution is also a cheaper and simpler method for preparing cultures from a single cell than optical tweezers and micromanipulators. We recommend having a water analysis done so that you know what constituents and at what concentration the sample is made of. Prepare the dilution of the test sample expected to contain between 30300 cfuml. Briefly, this method involves serial dilution of a bacterial suspension in sterile water blanks, which serve as a diluent of known volume.
We will extract 100 microliters of tetrahymena from stock to well. Procedure for bacteriophage plaque assay preparation of stock solution by serial dilution. Then, 1 ml from that mixture is added to 9 ml, and it is mixed together a 102 dilution. In microbiology, serial dilutions log dilutions are used to decrease a bacterial concentration to a required concentration for a specific test method, or to a.
Serial dilutions are often performed in steps of 10 or 100. I made this for my as biology students who dont take chemistry and needed some practice with serial dilutions before jumping straight into a full practical. The mpn procedure is a statistical method based upon the probability theory. In this test cardiolipin antigen is used as reagent to detect autoantibody in serum of patients. The first step in making a serial dilution is to take a known volume usually 1ml of stock and place it into a known volume of distilled water usually 9ml.
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