Method for treating living samples and analyzing the same
A method is provided by which a reagent is provided to a living sample, including contacting a thin-film containing a porous body with a living sample containing cells or tissues, and discharging a solution containing at least one or more reagents to a non-contact side of the thin-film by ink-jet to provide the solution to the specific area of the living sample through pores on the thin-film, wherein the smaller diameter of the pores on the porous body relative to the diameter of cells in the living sample enables the reagent to be provided to each individual cell separately. According to this method, the reagent and the reaction product can be fixed while maintaining positioning information of the target substance in the living sample, as a pretreatment for analyzing with high positioning accuracy the target substance in each individual cell in the living sample.
1. A method for providing a reagent to living cells in a sample comprising the steps of:
providing a film having a thickness between 1 μm and 10 μm comprising a porous body and having a top face and opposite bottom face, wherein pores of the porous body are not interconnected to each other and are present independently, wherein the pores have a diameter of 10 nm to 10 μm, and are smaller than a diameter of the cells in the sample, and wherein the pores perforate through the film from the top face to the bottom face such that the pores are vertical or have an angle of 15° or less from the vertical;
placing the living cells in the sample on a supporting member;
contacting the bottom face of the film directly with the living cells on the supporting member such that each pore of the porous body corresponds to one living cell on the supporting member; and
discharging by ink-jet a solution as a droplet containing at least one kind of reagent to the top face of the film not contacting the living cells such that the solution passes through the pores to the bottom face to contact the living cells to supply the reagent to each individual living cell separately.
2. The method for providing a reagent to living cells in a sample according to claim 1 , wherein the reagent is a compound that reacts chemically or biologically with a target substance in one or more of the living cells.
3. The method for providing a reagent to living cells in a sample according to claim 1 , further comprising a step of reading image information of the living cells to provide the reagent to each living cell of the sample by ink-jet in accordance with the image information.
4. A method for analyzing living cells in a sample, comprising the steps of:
providing to living cells in the sample a reagent using the method according to claim 1 ; wherein the reagent reacts with a target substance in the living cells; and
analyzing the living cells by microscopic observation or mass spectrometry, such that positioning information of the target substance is analyzed in each living cell.
5. The method for analyzing living cells in a sample according to claim 4 , wherein the mass spectrometry is time-of-flight secondary ion mass spectrometry (TOF-SIMS).