CN203150515U

CN203150515U - Adherent cell scanning electron microscope (SEM) carrier

Info

Publication number: CN203150515U
Application number: CN 201320043884
Authority: CN
Inventors: 宋智琦; 李乃浩
Original assignee: First Affiliated Hospital of Dalian Medical University
Current assignee: First Affiliated Hospital of Dalian Medical University
Priority date: 2013-01-28
Filing date: 2013-01-28
Publication date: 2013-08-21
Anticipated expiration: 2023-01-28

Abstract

The utility model discloses an adherent cell scanning electron microscope (SEM) carrier comprising a SEM sample holder (1); a top of the SEM sample holder (1) is fixed with a bottom plate (3) of a TC surface cell culture dish through an electric conduction adhesive layer (2); the bottom plate (3) of the TC surface cell culture dish is covered by a cell layer (4); an electric conduction film (5) is arranged on the cell layer (4); and a bus-bar (6) is in a bridge connection between the electric conduction film (5) and the electric conduction adhesive layer (2). Because the TC surface cell culture dish has very good endophilicity to the cells, so a coating layer is omitted to reduce cost; because side walls of the TC surface cell culture dish are finally cut off and the bottom plate covered by the cells is fixed on the sample holder, so cell surfaces are very clear, and phenomenon that cells are fixed on the sample holder with surfaces facing downwards will not happen; and the carrier is low in cost, can prevent cell form changes, and guarantees experiment effects.

Description

Attached cell ESEM carrier

Technical field

The utility model relates to a kind of attached cell ESEM carrier, and especially a kind of cost is low, avoid cellular morphology to change, guarantee the attached cell ESEM carrier of experiment effect.

Background technology

At present, at medical domain, the surface topography of observing attached cell with ESEM (electron microscope) is very general.Because ESEM is that the various physical signallings that utilize the fine focusing electron beam to eject when sample surfaces scans are modulated into picture, therefore attached cell must be prepared into can be for the carrier of scanning electron microscopic observation, i.e. attached cell ESEM carrier.Existing attached cell ESEM carrier is to be culture plate with the glass slide mostly, and its preparation method is that glass slide is put into Tissue Culture Dish basically behind ultraviolet disinfection, poly-D-lysine bag quilt, carries out cell and cultivates.After cell is cultured to desired density, add buffer solution in the culture dish cell is cleaned, and then with pH7.2 ~ 7.4, concentration is 2.5% glutaraldehyde fixed cell, after again cell being cleaned, with the ethanol of variable concentrations gradient to cell dehydration, drying and vacuumize processing.To be covered with cell and have the glass slide of bag tegillum to take out, and by conductive adhesive layer glass slide is fixed in the scanning electron microscope example holder, draw a bus from the conductive adhesive layer side, the other end of bus is placed on the cell face, with ion sputtering plated film instrument plating conductive layer, the other end of bus and conductive layer are fixedly connected.Because the cellular layer on the glass slide is colourless and very thin, therefore be difficult to distinguish cell face and glass slide bottom surface, occur sometimes the ventricumbent glass slide of cell is fixed on phenomenon in the sample holder.

TC superficial cell culture dish (Corning tissue-culture treated culture dishes) is existing cell culture apparatus ware, be with transparent, have the pure polystyrene manufacturing of certain toughness moral and form, inner surface covalent bond hydrogel layer, possess hydrophilic property and neutral charge, has the persistence of surface wettability and the stability of cell absorption, as special-purpose cell culture apparatus tool, cell there is good compatibility.Yet, up to now not about directly prepare the relevant report of attached cell ESEM carrier with TC superficial cell culture dish.

Summary of the invention

The utility model is in order to solve the above-mentioned technical problem of existing in prior technology, a kind of cost is low, avoid cellular morphology variation, assurance experiment effect attached cell ESEM carrier and preparation method to be provided.

Technical solution of the present utility model is: a kind of attached cell ESEM carrier, the scanning electron microscope example holder is arranged, the base plate that TC superficial cell culture dish is fixedly arranged by conductive adhesive layer in the scanning electron microscope example holder, base plate at TC superficial cell culture dish is covered with cellular layer, conducting film is arranged, bus cross-over connection conducting film and conductive adhesive layer on the cellular layer.

The utility model is directly to prepare attached cell ESEM carrier with TC superficial cell culture dish cultured cell and with the base plate of the TC superficial cell culture dish that is covered with cellular layer, because TC superficial cell culture dish has good compatibility to cell, therefore need not to wrap tegillum, reduced cost, owing to finally be that the base plate that will cut off TC superficial cell culture dish sidewall and be covered with cell is fixed in the sample holder, the cell face just comes into plain view, can not occur cell faced down is fixed on phenomenon in the sample holder, and it is low to have a cost, avoid cellular morphology to change, guarantee advantages such as experiment effect.

Description of drawings

Fig. 1 is the structural representation of the utility model embodiment.

Embodiment

The utility model carries out successively as follows:

A. cell is cultured to desired density in 35mm TC superficial cell culture dish;

B. using pH7.2 ~ 7.4, concentration is 2.5% glutaraldehyde fixed cell;

C. use the PBS buffer solution for cleaning cell 3 times of 0.1mol/ L, use the PBS buffer solution of 0.1mol/L to soak cell 12 ~ 15 hours again;

D. use 1 * PBS buffer solution for cleaning cell 1 time;

E. add the tert-butyl alcohol from low to high in cell according to concentration gradient, tert-butyl alcohol of every adding is is softly blown and beaten 3 ~ 5 times, leaves standstill 10 minutes, and 1 * PBS buffer solution for cleaning cell 1 time is until the last adding tert-butyl alcohol; Behind the last adding tert-butyl alcohol, just need not piping and druming, leave standstill, but directly under liquid nitrogen environment, make tert-butyl alcohol crystallization; The concentration gradient of the described tert-butyl alcohol is 10%, 25%, 50%, 60%, 70%, 80%, 90%, 100%;

F. TC superficial cell culture dish is inserted in the vacuum machine and vacuumize the tert-butyl alcohol of extraction crystallization etc.;

G. TC superficial cell culture dish sidewall is cut off, and the base plate that will be covered with the culture dish of cellular layer cuts to the required size of electron microscopic observation and shape and by conductive adhesive layer culture dish base plate (cellular layer up) is fixed in the scanning electron microscope example holder, draws a bus from the conductive adhesive layer side;

H. the other end with bus places on the cellular layer, with ion sputtering plated film instrument plating conductive layer, the other end of bus and conductive layer is fixedly connected.

The attached cell ESEM carrier of producing as shown in Figure 1, scanning electron microscope example holder 1 is arranged, the base plate 3 that TC superficial cell culture dish is fixedly arranged by conductive adhesive layer 2 in scanning electron microscope example holder 1, base plate 3 at TC superficial cell culture dish is covered with cellular layer 4, conducting film 5 is arranged on the cellular layer 4, and bus 6 cross-over connections are in conducting film 5 and conductive adhesive layer 2.

Claims

1. attached cell ESEM carrier, scanning electron microscope example holder (1) is arranged, it is characterized in that: the base plate (3) that TC superficial cell culture dish is fixedly arranged by conductive adhesive layer (2) in scanning electron microscope example holder (1), base plate (3) at TC superficial cell culture dish is covered with cellular layer (4), conducting film (5) is arranged, bus (6) cross-over connection conducting film (5) and conductive adhesive layer (2) on the cellular layer (4).