PRODUCTS
TI Blue
(Platinum blue)
An alternative to uranyl acetate for staining in TEM
It also enhances backscattered electron signals in SEM
335
solution 1ml x 1
335-1
Pack of 5 dry-hardened capsules, each of which is equal to 200l liquid.
335-2
Pack of 20 dry-hardened capsules,
each of which is equal to 50l liquid.
Comparison of TEM images obtained with TI Blue and conventional uranyl acetate
TI Blue + Lead acetate
rat liver
Uranyl acetate + Lead acetate
picture provider : Dr. Sumire Inaga, Department of Anatomy, Tottori University
TI Blue kit
Be useful anywhere and Anytime
The kit is supplied with a tray with 4 cavities with one containing dry-hardened TI Blue (see the image below). Of rest three cavities, one is for fixative (0.1M phosphate or cacodylate buffer containing 2.5% glutaraldehyde) and two others are for distilled water for rinsing after fixing or staining. Choice of cavities for fixative and distilled water is left to the users.
Low vacuum SEM for paraffin sections
After staining with TI Blue, paraffin sections mounted onto a slide can be examined directly in SEM using a variable pressure, or low vacuum, mode without need of deposition of a thin metallic coating to avoid sample surfacecharging.*
LM and LVSEM images of the same paraffin section from the rat tongue stained with TI Blue. Red arrows indicate
Mast cells by LM and LVSEM
*reference ;
Low vacuum scanning electron microscopy for paraffin sections utilizing the differential stainability of cells and tissues with platinum blue
S.Inaga, S.Hirashima, K.Tanaka, T.Katsumoto, T.Kameie, H.Nakane, T.Nagumo,
Arch.Histol Cytol, 72 (2):101-106(2009)
How to use TI Blue
Adjust pH immediately before use.
After pH adjustment, use up the solution in a day.
TI Blue Kit
The kit is supplied with a tray with 4 cavities with one containing dry-haedened TI Blue (see the picture below). Of rest three cavities, one is for fixative (0.1M phosphate or cacodylate buffer containing 2.5% glutaraldehyde) and two others are for distilled water for rinsing after fixing or staining.
Choice of cavities for fixative and distilled water is left to the users.
Procedure
Fill one of three empty cavities with fixative and two others with distilled water for rinsing.
Take out all solution for dilution of TI Blue(`200 l, pH 9.2) from the tube using micro-pipet or syringe and pour it into the cavity containing TI Blue.
(Make sure TI Blue is completely dissolved before use)
Immerse the sample into the fixative, take it out and put it into the cavity filled with distilled water for 1`2min.
Then, take the sample out from distilled water, remove residual distilledwater using a filter paper in the Kit before immersion into the TI Blue stain solution for 15 ` 20min.
(time required for staining depends on the nature of sample)
After staining, take out the sample from the stain solution, put it into the cavity filled with fresh distilled water for 1 ` 2 min for rinsing.
After rinsing, take out the sample, remove residual distilled water by filter paper and set it to the SEM specimen holder.
Examination in the BSE imaging mode at low vacuum condition is recommended.
TI Blue Small Kit
This reagent is mainly useful for BSE imaging mode at low vacuum condition.
The kit contains 20 tubes containing dry-hardened TI Blue at the bottom of each tube.
The kit also contains a tube which contains 2ml of solution (pH9.2) for dilution.
To use ; take 50l of solution from the attached tube by using micro-pipet or syringe and pour it into one of the tubes with TI Blue.
(Make sure TI Blue is dissolved completely)
Where required you can adjust concentration by changing the amount of solution.
TI Blue solution
TI Blue is supplied as 1 ml, concentrated solution kept in a bottle with a cap which central part is made of silicone to allow easy take out of solution using a syringe with a filter.
It is used after appropriate dilution.
Procedure
Staining of ultra-thin sections for TEM examination
Staining procedure is essentially the same as for uranyl acetate.
To find an optimum concentration for your specimen, start from a solution diluted by 10x in distilled water or in 50% methanol with 10 min staining time. See whether your specimen is under-stained or over-stained. Then, change the concentration of stain solution and/or staining time accordingly. Post-staining with lead citrate is recommended to get better results.
For negative staining, use the stain solution as supplied or after appropriate dilution (for some specimens, TI Blue may not work effectively).
Staining for SEM examination
Use the TI Blue solution as supplied and adjust the pH of solution to 9 or above through addition of NH4OH solution (pH of TI Blue concentrated solution is pH 3).
Staining will take place in 15 `20 min at room temperature depending on the specimen.
After staining, rinse the specimen in distilled water for 1 ` 2 min, remove residual water by filter paper. The wet sample is now ready for examination in SEM.; low vacuum, BSE imaging mode is recommended.
Low vaccume SEM for paraffin sections*
The section on the slides are deparaffinized with xylene and given an alcohol descending series then transferred to distilled water.
Remove distilled water and pour TI Blue solution onto section then leave it for 15 to 20 minutes at room temperature.
After staining, rinse the slide with specimen in distilled water for 1 to 2 minutes.
Remove distilled water from the slide and fix it onto the specimen holder of SEM with carbon adhesive tape before introducing into the specimen chamber of SEM.
*reference ;Rapid three-dimensional analysis of renal biopsy sections by low vacuum scanning electron microscopy
S.Inaga, M.Kato, S.Hirashima, C.Munemura, S.Okada, T.Kameie, T.Katsumoto, H.Nakane, K.Tanaka, K.Hayashi, T.Naguro
Arch.Histol Cytol, vol.73,(3):113-125(2010/2011)
On disposal of waste solution, please follow the regulation for the hazardous waste containing heavy metals.
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