07.100.01 (Microbiology in general) 标准查询与下载

ASTM E2186-02 用彗星分析法测定真核细胞中DNA单线损伤的标准指南

1.1 This guide covers the recommended criteria for performing a single-cell gel electrophoresis assay (SCG) or Comet assay for the measurement of DNA single-strand breaks in eukaryotic cells. The Comet assay is a very sensitive method for detecting strand breaks in the DNA of individual cells. The majority of studies utilizing the Comet assay have focused on medical applications and have therefore examined DNA damage in mammalian cells in vitro and in vivo (1-4). There is increasing interest in applying this assay to DNA damage in freshwater and marine organisms to explore the environmental implications of DNA damage.1.1.1 The Comet assay has been used to screen the genotoxicity of a variety of compounds on cells in vitro and in vivo (5-7), as well as to evaluate the dose-dependent anti-oxidant (protective) properties of various compounds (3, 8-11). Using this method, significantly elevated levels of DNA damage have been reported in cells collected from organisms at polluted sites compared to reference sites (12-15). Studies have also found that increases in cellular DNA damage correspond with higher order effects such as decreased growth, survival, and development, and correlate with significant increases in contaminant body burdens (13, 16).1.2 This guide presents protocols that facilitate the expression of DNA alkaline labile single-strand breaks and the determination of their abundance relative to control or reference cells. The guide is a general one meant to familiarize lab personnel with the basic requirements and considerations necessary to perform the Comet assay. It does not contain procedures for available variants of this assay, which allow the determination of non-alkaline labile single-strand breaks or double-stranded DNA strand breaks (8), distinction between different cell types (13), identification of cells undergoing apoptosis (programmed cell death, (1, 17)), measurement of cellular DNA repair rates (10), detection of the presence of photoactive DNA damaging compounds (14), or detection of specific DNA lesions (3, 18).1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory requirements prior to use.1.3 This guide is arranged as follows: SectionScope1Referenced Documents2Terminology3Summary of Guide4Significance and Use5Equipment and Reagents6Assay Procedures7Treatment of Data8Reporting Data9Keywords10Annex A1References

Standard Guide for Determining DNA Single-Strand Damage in Eukaryotic Cells Using the Comet Assay

ASTM E2197-02 液体化学杀菌剂的杀菌,杀病毒剂的,杀细菌的,杀分枝杆菌的,杀芽孢菌剂活力测定的标准数量盘载波试验方法

1.1 The method is designed to evaluate the ability of liquid chemical germicides to inactivate vegetative bacteria, viruses, fungi, mycobacteria and bacterial spores in the presence of a soil load (1,2) on disk carriers that represent environmental surfaces and medical devices. It is also designed to have survivors that can be compared to mean of no less than three control carriers to determine if the performance standard has been met. For proper statistical evaluation of the results, the size of the test inoculum should be sufficiently large to take into account both the performance standard and the experimental variation in the results.1.2 The test protocol does not include any wiping or rubbing action. It is, therefore, not designed for testing germicide-soaked wipes.1.3 This test method should be performed by persons with training in microbiology in facilities designed and equipped for work with infectious agents at the appropriate biosafety level ().1.4 In this test method, metric units are used for all applications, except for distance in which case inches are used and metric units follow.1.5 It is the responsibility of the investigator to determine whether Good Laboratory Practice Regulations (GLPs) are required and to follow them where appropriate (40 CFR, Part 160 for EPA submissions and 21 CFR, Part 58 for FDA submissions).1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

Standard Quantitative Disk Carrier Test Method for Determining the Bactericidal, Virucidal, Fungicidal, Mycobactericidal and Sporicidal Activities of Liquid Chemical Germicides

ASTM E1493-01 抗菌素M13或其DNA识别的标准导则

1.1 This guide covers the identification of bacteriophage M13 used in biotechnology. 1.2 There are many variants of M13 that have been developed specifically for cloning technology. These variants have foreign DNA inserted into the M13 genome, causing the M13 to differ in size and genotype. 1.3 If the M13 is to be used to construct a recombinant molecule, then the criteria described in Section 6 should be used to characterize the newly made DNA.

Standard Guide for Identification of Bacteriophage M13 or Its DNA

ASTM E2180-01 聚合或疏水材料中掺入的杀微生物剂活性测定的标准试验方法

This method can be used to evaluate effectiveness of incorporated/bound antimicrobials in hydrophobic materials such as plastics, epoxy resins, as well as other hard surfaces. The aqueous based bacterial inoculum remains in close, uniform contact in a “pseudo-biofilm” state with the treated material. The percent reduction in the surviving populations of challenge bacterial cells at 24 h versus those recovered from a non-treated control is determined. The hydrophobic substrate may be repeatedly tested over time for assessment of persistent antimicrobial activity.1.1 This test method is designed to evaluate (quantitatively) the antimicrobial effectiveness of agents incorporated or bound into or onto mainly flat (two dimensional) hydrophobic or polymeric surfaces. The method focuses primarily on assessing antibacterial activity; however, other microorganisms such as yeast and fungal conidia may be tested using this method.1.2 The vehicle for the inoculum is an agar slurry which reduces the surface tension of the saline inoculum carrier and allows formation of a "pseudo-biofilm," providing more even contact of the inoculum with the test surface.1.3 This method can confirm the presence of antimicrobial activity in plastics or hydrophobic surfaces and allows determination of quantitative differences in antimicrobial activity between untreated plastics or polymers and those with bound or incorporated low water-soluble antimicrobial agents. Comparisons between the numbers of survivors on preservative-treated and control hydrophobic surfaces may also be made.1.4 The procedure also permits determination of "shelf-life" or long term stability of an antimicrobial treatment which may be achieved through testing both non-washed and washed samples over a time span.1.5 Knowledge of microbiological techniques is required for these procedures.1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

Standard Test Method for Determining the Activity of Incorporated Antimicrobial Agent(s) In Polymeric or Hydrophobic Materials

ASTM F2149-01 细胞自动分析的标准试验方法.单细胞悬浮液计数和分级的电敏域法

1.1 This test method, provided the limitations are understood, covers a procedure for both the enumeration and measurement of size distribution of most all cell types. The instrumentation allows for user-selectable cell size settings, hence, this test method is not restricted to specific cell types. The method is appropriate for suspension as well as adherent cell cultures (1). This is a quantitative laboratory method not intended for on-line or field use. Results may be reported as number of cells per millilitre or total number of cells per volume of cell suspension analyzed. Both count and size distribution may be expressed in cell micron diameter or volume, femtolitres.1.2 Cells commonly used in tissue-engineered medical products (2) routinely are analyzed. Examples are chondrocytes (3), fibroblasts (4), and keratinocytes (5). Szabo et al used the method for both pancreatic islet number and volume measurements (6). In addition, instrumentation using the electrical sensing zone technology was used for both count and size distribution analyses of porcine hepatocytes placed into hollow fiber cartridge extracorporeal liver assist systems. In this study (7), and others (6, 8), the automated electrical sensing zone method was clearly validated for superior accuracy and precision when compared to the conventional manual method, visual cell counting under a microscope using a hemocytometer. This validation has been demonstrated over a wide variety of cell types. In addition, the automated procedure is rapid, rugged, and cost effective; it also minimizes operator-to-operator variability inherent in manual techniques.1.3 This instrumentation is manufactured by a variety of companies; however, the principle used in all is electrical impedance. This test method, for cell counting and sizing, is based on the detection and measurement of changes in electrical resistance produced by a cell, suspended in a conductive liquid, traversing through a small aperture (see (Fig. 1)). When cells are suspended in a conductive liquid, phosphate-buffered saline for instance, they function as discrete insulators. When the cell suspension is drawn through a small cylindrical aperture, the passage of each cell changes the impedance of the electrical path between two submerged electrodes located on each side of the aperture. An electrical pulse, suitable for both counting and sizing, results from the passage of each cell through the aperture. The path through the aperture, in which the cell is detected, is known as the "electronic sensing zone." This test method permits the selective counting of cells within very narrow size distribution ranges by electronic selection of the generated pulses. While the number of pulses indicates cell count, the amplitude of the electrical pulse produced depends on the cell''s volume. The baseline resistance between the electrodes is due to the resistance of the conductive liquid within the boundaries of the aperture. The presence of cells within the "electronic sensing zone" raises the resistance of the conductive pathway that depends on the volume of the cell. Analyses of the behavior of cells within the aperture demonstrates that the height of the pulse produced by the cell is the parameter that most nearly shows proportionality to the cell volume.1.4 Limitations are discussed as follows:1.4.1 Coincidence8212;Occasionally, more than a single cell transverses the aperture simultaneously. Only a single larger pulse, as opposed to two individual pulses, is generated. The result is a lower cell count and higher cell volume measurement. The frequency of coincidence is a statistically predictable function of cell concentration that is corrected by the instrument. This is called coincidence correction (8). This phenomenon may be minimized, thus ensuring greater result accuracy, by using relatively low cell ......

Standard Test Method for Automated Analyses of Cells8212;the Electrical Sensing Zone Method of Enumerating and Sizing Single Cell Suspensions

ASTM F2149-01(2007) 细胞自动分析的标准试验方法.单细胞悬浮液计数和分级的电敏域法

This assay is used in university tissue culture laboratories, government research, and hospital, biomedical, and pharmaceutical laboratories to automate cell counting and sizing. This instrumentation provides very rapid, accurate, and precise results for any tissue culture facility. In addition, as noted, since the cell sizes to be analyzed by the instrument are set by the user, the analyses may be done on virtually any species of cells and cell type; it is not restricted to human cells or blood cells. The electrical sensing zone methodology was introduced in the mid 1950s (9). Since this time, there have been substantial improvements which have enhanced the operatorrsquo;ease of use. Among these are the elimination of the mercury manometer, reduced size, greater automation, and availability of comprehensive statistical computer programs. This instrumentation offers a rapid result as contrasted to the manual counting of cells using the standard counting chamber, hemocytometer. The counting chamber is known to have an error of 10 to 30 %, as well as being very time consuming (10). In addition, when counting and sizing porcine hepatocytes, Stegemann et al concluded that the automated, electrical sensing zone method provided significantly greater accuracy, precision, and speed, for both counts and size, compared to the conventional microscopic or the cell mass-based method (7).1.1 This test method, provided the limitations are understood, covers a procedure for both the enumeration and measurement of size distribution of most all cell types. The instrumentation allows for user-selectable cell size settings, hence, this test method is not restricted to specific cell types. The method is appropriate for suspension as well as adherent cell cultures (). This is a quantitative laboratory method not intended for on-line or field use. Results may be reported as number of cells per millilitre or total number of cells per volume of cell suspension analyzed. Both count and size distribution may be expressed in cell micron diameter or volume, femtolitres.1.2 Cells commonly used in tissue-engineered medical products () routinely are analyzed. Examples are chondrocytes (), fibroblasts (), and keratinocytes (). Szabo et al used the method for both pancreatic islet number and volume measurements (). In addition, instrumentation using the electrical sensing zone technology was used for both count and size distribution analyses of porcine hepatocytes placed into hollow fiber cartridge extracorporeal liver assist systems. In this study (), and others (, ), the automated electrical sensing zone method was clearly validated for superior accuracy and precision when compared to the conventional manual method, visual cell counting under a microscope using a hemocytometer. This validation has been demonstrated over a wide variety of cell types. In addition, the automated procedure is rapid, rugged, and cost effective; it also minimizes operator-to-operator variability inherent in manual techniques.1.3 This instrumentation is manufactured by a variety of companies; however, the principle used in all is electrical impedance. This test method, for cell counting and sizing, is based on the detection and measurement of changes in electrical resistance produced by a cell, suspended in a conductive liquid, traversing through a small aperture (see ()). When cells are suspended in a conductive liquid, phosphate-buffered saline for instance, they function as discrete insulators. When the cell suspension is drawn through a small cylindrical aperture, the passage of each cell changes the impedance of the electrical path between two submerged electrodes located on each side of the aperture. ......

Standard Test Method for Automated Analyses of Cells-the Electrical Sensing Zone Method of Enumerating and Sizing Single Cell Suspensions

ASTM E1531-00 用琼脂生长法直接检测细胞培养中枝原体属染菌的标准实施规程

1.1 This practice covers the procedures used for detection of mycoplasma contamination by direct microbiological culture.1.2 This practice does not cover indirect methods for detection of mycoplasma such as DNA staining, biochemical detection, or genetic probes.1.3 This practice does not cover methods for identification of mycoplasma organisms.1.4 This practice will not detect cultivar strains (1) of Mycoplasma hyorhinis.1.5 This practice is not intended for use in detection of mycoplasma contamination in sera, culture media, vaccines, or other systems.1.6 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

Standard Practice for Detection of Mycoplasma Contamination of Cell Cultures by Growth on Agarose Medium

ASTM D5245-92(1998) 微生物分析用清洁的实验室玻璃器皿、塑料器具和设备的标准实施规程

1.1 In microbiology, clean glassware is crucial to ensure valid results. Previously used or new glassware must be thoroughly cleaned. Laboratory ware and equipment that are not chemically clean are responsible for considerable losses in personnel time and supplies in many laboratories. These losses may occur as down time when experiments clearly have been adversely affected and as invalid data that are often attributed to experimental error. Chemical contaminants that adversely affect experimental results are not always easily detected. This practice describes the procedures for producing chemically clean glassware. 1.2 The values stated in SI units are to be regarded as the standard. 1.3 This practice does not purport to address all of the safety problems, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. For specific precautions, see Section 5, 7.3.1, and Notes 1 and 2.

Standard Practice for Cleaning Laboratory Glassware, Plasticware, and Equipment Used in Microbiological Analyses

ASTM D5245-92(2012) 微生物分析用清洁的实验室玻璃器皿、塑料器具和设备的标准实施规程

This practice provides uniform guidance for cleaning the laboratory glassware, plasticware, and equipment used in routine microbiological analyses. However, tests that are extremely sensitive to toxic agents (such as virus assays) may require more stringent cleaning practices. 1.1 In microbiology, clean glassware is crucial to ensure valid results. Previously used or new glassware must be thoroughly cleaned. Laboratory ware and equipment that are not chemically clean are responsible for considerable losses in personnel time and supplies in many laboratories. These losses may occur as down time when experiments clearly have been adversely affected and as invalid data that are often attributed to experimental error. Chemical contaminants that adversely affect experimental results are not always easily detected. This practice describes the procedures for producing chemically clean glassware. 1.2 The values stated in SI units are to be regarded as the standard. 1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use. For specific precautions, see Section 5, 7.3.1, and Note 1 and Note 2.

Standard Practice for Cleaning Laboratory Glassware, Plasticware, and Equipment Used in Microbiological Analyses

ASTM E1202-87(2003) 微核检测标准制定用标准指南

Micronucleus assays for genetic damage have been developed in many types of eucaryotic cells, both in vitro and in vivo. The occurrence of micronuclei is indicative of chromosomal damage or mitotic spindle dysfunction.1.1 This guide covers minimal criteria which should be met by a micronucleus assay system prior to the development of an ASTM Standard or Guide for the conduct of that assay.1.2 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

Standard Guide for Development of Micronucleus Assay Standards

ASTM E1202-87(1998) 微核检测标准制定用标准指南

1.1 This guide covers minimal criteria which should be met by a micronucleus assay system prior to the development of an ASTM Standard or Guide for the conduct of that assay. 1.2 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

Standard Guide for Development of Micronucleus Assay Standards