An agar plate is a Petri dish that contains a growth medium solidified with agar, used to culture microorganisms. Sometimes selective compounds are added to influence growth, such as antibiotics.
Replica plating is a microbiological technique in which one or more secondary Petri plates containing different solid (agar-based) selective growth media are inoculated with the same colonies of microorganisms from a primary plate, reproducing the original spatial pattern of colonies. The technique involves pressing a velveteen-covered disk, and then imprinting secondary plates with cells in colonies removed from the original plate by the material. Generally, large numbers of colonies are replica plated due to the difficulty in streaking each out individually onto a separate plate.
A microbiological culture, or microbial culture, is a method of multiplying microbial organisms by letting them reproduce in predetermined culture medium under controlled laboratory conditions. Microbial cultures are foundational and basic diagnostic methods used as research tools in molecular biology.
Bacteriological water analysis is a method of analysing water to estimate the numbers of bacteria present and, if needed, to find out what sort of bacteria they are. It represents one aspect of water quality. It is a microbiological analytical procedure which uses samples of water and from these samples determines the concentration of bacteria. It is then possible to draw inferences about the suitability of the water for use from these concentrations. This process is used, for example, to routinely confirm that water is safe for human consumption or that bathing and recreational waters are safe to use.
The Miles and Misra Method is a technique used in Microbiology to determine the number of colony forming units in a bacterial suspension or homogenate. The technique was first described in 1938 by Miles, Misra and Irwin who at the time were working at the LSHTM. The Miles and Misra method has been shown to be precise.
In microbiology, streaking is a technique used to isolate a pure strain from a single species of microorganism, often bacteria. Samples can then be taken from the resulting colonies and a microbiological culture can be grown on a new plate so that the organism can be identified, studied, or tested.
Total viable organism is a term used in microbiology to quantify the amount of microorganisms present in a sample. Each sample is usually cultured on a variety of agar plates often containing different types of selective media. The colony-forming units (CFUs) are calculated after allowing time for growth.
The Neogen Petrifilm plate is an all-in-one plating system made by the Food Safety Division of the Neogen Corporation. They are heavily used in many microbiology-related industries and fields to culture various micro-organisms and are meant to be a more efficient method for detection and enumeration compared to conventional plating techniques. A majority of its use is for the testing of foodstuffs.
The minimum bactericidal concentration (MBC) is the lowest concentration of an antibacterial agent required to kill a particular bacterium. It can be determined from broth dilution minimum inhibitory concentration (MIC) tests by subculturing to agar plates that do not contain the test agent. The MBC is identified by determining the lowest concentration of antibacterial agent that reduces the viability of the initial bacterial inoculum by ≥99.9%. The MBC is complementary to the MIC; whereas the MIC test demonstrates the lowest level of antimicrobial agent that inhibits growth, the MBC demonstrates the lowest level of antimicrobial agent that results in microbial death. This means that even if a particular MIC shows inhibition, plating the bacteria onto agar might still result in organism proliferation because the antimicrobial did not cause death. Antibacterial agents are usually regarded as bactericidal if the MBC is no more than four times the MIC. Because the MBC test uses colony-forming units as a proxy measure of bacterial viability, it can be confounded by antibacterial agents which cause aggregation of bacterial cells. Examples of antibacterial agents which do this include flavonoids and peptides.
Total viable count (TVC), gives a quantitative estimate of the concentration of microorganisms such as bacteria, yeast or mould spores in a sample. The count represents the number of colony forming units (cfu) per g of the sample.
Plate count agar (PCA), also called standard methods agar (SMA), is a microbiological growth medium commonly used to assess or to monitor "total" or viable bacterial growth of a sample. PCA is not a selective medium.
Transformation efficiency refers to the ability of a cell to take up and incorporate exogenous DNA, such as plasmids, during a process called transformation. The efficiency of transformation is typically measured as the number of transformants per microgram of DNA added to the cells. A higher transformation efficiency means that more cells are able to take up the DNA, and a lower efficiency means that fewer cells are able to do so.
Cell counting is any of various methods for the counting or similar quantification of cells in the life sciences, including medical diagnosis and treatment. It is an important subset of cytometry, with applications in research and clinical practice. For example, the complete blood count can help a physician to determine why a patient feels unwell and what to do to help. Cell counts within liquid media are usually expressed as a number of cells per unit of volume, thus expressing a concentration.
Virtual colony count (VCC) is a kinetic, 96-well microbiological assay originally developed to measure the activity of defensins. It has since been applied to other antimicrobial peptides including LL-37. It utilizes a method of enumerating bacteria called quantitative growth kinetics, which compares the time taken for a bacterial batch culture to reach a threshold optical density with that of a series of calibration curves. The name VCC has also been used to describe the application of quantitative growth kinetics to enumerate bacteria in cell culture infection models. Antimicrobial susceptibility testing (AST) can be done on 96-well plates by diluting the antimicrobial agent at varying concentrations in broth inoculated with bacteria and measuring the minimum inhibitory concentration that results in no growth. However, these methods cannot be used to study some membrane-active antimicrobial peptides, which are inhibited by the broth itself. The virtual colony count procedure takes advantage of this fact by first exposing bacterial cells to the active antimicrobial agent in a low-salt buffer for two hours, then simultaneously inhibiting antimicrobial activity and inducing exponential growth by adding broth. The growth kinetics of surviving cells can then be monitored using a temperature-controlled plate reader. The time taken for each growth curve to reach a threshold change in optical density is then converted into virtual survival values, which serve as a measure of antimicrobial activity.
Spot analysis, spot test analysis, or spot test is a chemical test, a simple and efficient technique where analytic assays are executed in only one, or a few drops, of a chemical solution, preferably in a great piece of filter paper, without using any sophisticated instrumentation. The development and popularization of the test is credited to Fritz Feigl.
A spiral plater is an instrument used to dispense a liquid sample onto a Petri dish in a spiral pattern. Commonly used as part of a CFU count procedure for the purpose of determining the number of microbes in the sample. In this setting, after spiral plating, the Petri dish is incubated for several hours after which the number of colony forming microbes (CFU) is determined. Spiral platers are also used for research, clinical diagnostics and as a method for covering a Petri dish with bacteria before placing antibiotic discs for AST.
Agar dilution is one of two methods used by researchers to determine the minimum inhibitory concentration (MIC) of antibiotics. It is the dilution method most frequently used to test the effectiveness of new antibiotics when a few antibiotics are tested against a large panel of different bacteria.
Impedance microbiology is a microbiological technique used to measure the microbial number density of a sample by monitoring the electrical parameters of the growth medium. The ability of microbial metabolism to change the electrical conductivity of the growth medium was discovered by Stewart and further studied by other scientists such as Oker-Blom, Parson and Allison in the first half of 20th century. However, it was only in the late 1970s that, thanks to computer-controlled systems used to monitor impedance, the technique showed its full potential, as discussed in the works of Fistenberg-Eden & Eden, Ur & Brown and Cady.
In microbiology, the term isolation refers to the separation of a strain from a natural, mixed population of living microbes, as present in the environment, for example in water or soil, or from living beings with skin flora, oral flora or gut flora, in order to identify the microbe(s) of interest. Historically, the laboratory techniques of isolation first developed in the field of bacteriology and parasitology, before those in virology during the 20th century.
In microbiology, colonial morphology refers to the visual appearance of bacterial or fungal colonies on an agar plate. Examining colonial morphology is the first step in the identification of an unknown microbe. The systematic assessment of the colonies' appearance, focusing on aspects like size, shape, colour, opacity, and consistency, provides clues to the identity of the organism, allowing microbiologists to select appropriate tests to provide a definitive identification.
