GB/T 35098-2018 in English

VALID

Microbeam analysis—Transmission electron microscopy—Morphological identification method of plant virusesby transmission electron microscopy

Issued on:2018-05-14
Implemented on:2019-04-01
File Format:PDF
Delivery:Via email within 1~3 business days

Price(USD)： $250.00
$243.00

Standard No:	GB/T 35098-2018
Document status:	VALID
Title in English:	Microbeam analysis—Transmission electron microscopy—Morphological identification method of plant virusesby transmission electron microscopy
Title in Chinese:	微束分析　透射电子显微术　植物病毒形态学的透射电子显微镜鉴定方法
Language:	English
File Format:	Electronic (PDF)
Delivery:	Via email within 1~3 business days
Issued on:	2018-05-14
Implemented on:	2019-04-01
ICS Classification:	71.040.40-Chemical analysis
Chinese Classification:	G04-Basic standards and general methods
Professional Classification:	GB-National Standard
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《GB/T 35098-2018微束分析透射电子显微术植物病毒形态学的透射电子显微镜鉴定方法》由TC38（全国微束分析标准化技术委员会）归口，主管部门为国家标准化管理委员会。

Introduction

Analysis of the standard technical framework

This standard systematically standardizes the three core technologies for identifying plant viruses using transmission electron microscopy: negative staining, immunoelectron microscopy and ultrathin sectioning technology. The technical framework is shown in the figure below:

Technical method	Resolution	Applicable scenarios	Preparation time
Negative staining method	2-5nm	Rapid diagnosis of virus suspension	15-30min
Immunoelectron microscopy	5-10nm	Virus serological identification	2-4h
Ultrathin section	1-2nm	Intracellular virus localization	3-5 days

Core technology innovations

1. Composite staining system

The three types of negative staining solutions recommended by the standard are complementary:

Phosphotungstic acid (PTA): a universal stain with good contrast for most viruses at pH 6.7
Uranium acetate: high-resolution staining (pH 4.2), but salt interference should be avoided
Ammonium molybdate: specifically for enveloped viruses (pH 7.0), with minimal destructiveness

2. Immunoenrichment technology

The immunoadsorption-modification method achieves virus enrichment through a three-step reaction:

The grid is pre-coated with specific antibodies
Viral antigen-antibody complex formation
Secondary antibody labeling enhances the signal

Key operating specifications

Ultrathin section quality control

The standard clearly stipulates that the section thickness should be controlled in the range of 50-100nm and through:

Double fixation: glutaraldehyde (2-3%) + osmium tetroxide (1%)
Gradient dehydration: ethanol concentration increases from 50% to 100%
Spurr resin embedding: low viscosity formula ensures penetration effect

Electron microscopy observation parameters

Observation mode	Accelerating voltage	Magnification	Scale requirements
Negative staining	60-80kV	4,000-100,000×	Must include
Ultrathin sectioning	80-100kV	1,000-50,000×	Must include

Recommendations for implementation of the standard

Laboratory configuration plan

According to clause 5.1 of the standard, the basic laboratory should be equipped with:

Transmission electron microscope (including CCD camera)
Ultrathin sectioner + diamond knife
Glow discharge instrument (grid hydrophilization treatment)
Constant temperature and humidity system (temperature 20±5℃, humidity <60%)

Construction of virus morphology database

It is recommended to refer to Appendix B to establish a localized database, including:

Typical virus morphology of 118 genera in 26 families
Characteristic inclusion body structure
Atlas of cytopathological changes

More samples: YD/T 3934-2021 JJG 21-2008 JGJ 94-2008

Sample only — not a preview of GB/T 35098-2018

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