Method for collecting and preparing specimens for immune reactions

Claim 1

A method for collecting particles of an immunoreactive specimen for electron microscopy examination, comprising the steps of: (a) fixing antibody particles to non-magnetic particles having a mass greater than said specific antibody particles to form carrier fixed particles; (b) subjecting a specimen and said carrier fixed particles to an immunoreaction to form reacted immunocomplex particles; (c) labeling said reacted immunocomplex particles with magnetic micro-particles having a particle size of 10 to 30 nm; (d) separating and removing unreacted particles of said antibody by centrifugation; and (e) performing local concentration of the immunocomplex to a predetermined region by applying an external gradient magnetic field, which is produced by an electromagnet and a permanent magnetic pole; (f) washing to remove extraneous particles and (g) retrieving the immunocomplex by direct magnetic adsorption towards the magnetic pole at said region of local concentration.

Claim 2

A method for collecting particles of an immunoreactive specimen for electron microscopy examination, comprising the steps of: (a) fixing antibody particles to non-magnetic particles having a mass greater than said specific antibody particles to form carrier fixed particles; (b) subjecting a specimen and said carrier fixed particles to an immunoreaction to form reacted immunocomplex particles; (c) labeling said reacted immunocomplex particles with magnetic micro-particles having a particle size of 10 to 30 nm; (d) separating and removing unreacted particles of said antibody by centrifugation; and (e) performing local concentration of the immunocomplex to a predetermined region by applying an external gradient magnetic field, which is produced by an electromagnet and a permanent magnetic pole; (f) washing to remove extraneous particles and (g) retrieving the immunocomplex by direct magnetic adsorption towards the magnetic pole at said region of local concentration.

Claim 9

A method for preparing a specimen, comprising injecting a specimen to a magnetic-labeled body holding position in a reactor vessel at which a magnetic-labeled antibody comprising a magnetic micro-particle having a particle size of 10 to 30 nm and an antibody or antigen bound to the magnetic micro-particle is held localized by means of an external magnetic force, to subject the specimen and the magnetic-labeled body to an antigen-antibody reaction, washing to remove extraneous particles, recovering the magnetic-labeled body by releasing the application of the external magnetic force, and holding the magnetic-labeled body by magnetizing a magnetic member inserted in the inside of the reactor vessel, and recovering the magnetic-labeled specimen by dipping said magnetic member entirely in a washing solution and demagnetizing the magnetic member, followed by vibrating said magnetic member so as to release said specimen in said washing solution.

Claim 10

A method for preparing a specimen, comprising injecting a specimen to a magnetic-labeled body holding position in a reactor vessel at which a magnetic-labeled antibody comprising a magnetic micro-particle having a particle size of 10 to 30 nm and an antibody or antigen bound to the magnetic micro-particle is held localized by means of an external magnetic force, to subject the specimen and the magnetic-labeled body to an antigen-antibody reaction, washing to remove extraneous particles, recovering the magnetic-labeled body by releasing the application of the external magnetic force, and holding the magnetic-labeled body by magnetizing a magnetic member inserted in the inside of the reactor vessel, and recovering the magnetic-labeled specimen by dipping said magnetic member entirely in a washing solution and demagnetizing the magnetic member, followed by vibrating said magnetic member so as to release said specimen in said washing solution.

Claim 11

A method for preparing an electron microscopy specimen, comprising the steps of: preparing a suspension of a magnetically labeled analyte specimen particles containing target antigen or antibody particles; applying an external gradient magnetic field to said suspension so as to concentrate the magnetic labeled analyte particles in a localized region of said suspension; inserting a grid for use in electron microscopy in said localized region of said suspension; applying an auxiliary magnetic field on said grid so as to adsorb said analyte particles on a surface of said grid; subjecting said analyte particles magnetically adsorbed on said grid surface to repeated washing cycles with purified water, while applying a magnetic force, so as to maintain analyte particles consisting essentially of target antigen or antibody particles on said grid surface; and turning-off said gradient magnetic field and removing said grid for electron microscope examination of target antigen particles.

Claim 12

A method for preparing an electron microscopy specimen, comprising the steps of: preparing a suspension of a magnetically labeled analyte specimen particles containing target antigen or antibody particles; applying an external gradient magnetic field to said suspension so as to concentrate the magnetic labeled analyte particles in a localized region of said suspension; inserting a grid for use in electron microscopy in said localized region of said suspension; applying an auxiliary magnetic field on said grid so as to adsorb said analyte particles on a surface of said grid; subjecting said analyte particles magnetically adsorbed on said grid surface to repeated washing cycles with purified water, while applying a magnetic force, so as to maintain analyte particles consisting essentially of target antigen or antibody particles on said grid surface; and turning-off said gradient magnetic field and removing said grid for electron microscope examination of target antigen particles.