Description

[0001] The invention relates to a process for the identification and individualization of samples, particularly of cell and tissue samples, by an in particular blockshaped, on a microtome or ultramicrotome cuttable, coded material. Especially agar may be suited as such material.

[0002] Particularly in the field of human pathology, the structure of cell and tissue samples is examined by light microscopy. Until the final examination under the microscope the samples are processed in several steps (including e.g. the tissue sampling and removal out of the body by a clinical physician, the transport of the sample in a container to the institute of pathology, the unbagging of the sample from the container for macroscopic evaluation, the transfer of the sample into a plastic cassette for paraffinization, the installment of the sample into a steel embedding mold for the construction of a paraffin block, the mounting of a section onto a slide), at which the samples are stored in different containers and sample holders (e.g. slides). So far, only the containers or the sample holders are marked and thereby individualized (e.g. by numbers), but not the samples theirselves. For this reason, sample mix-ups can easily happen at the different processing steps where a manual transfer of the samples e.g. from one container to another is performed, and are hardly to detect.

[0003] By the present invention, however, the sample material is uniquely marked

immediately after the removal from the body by pouring the specimens together with coded agar bodies into agar blocks or by processing coded agar bodies together with the specimens in a sealed container like the Tissue-Tek® Paraform® Sectionable Cassette System

(SAKURA FINETEK USA, Inc., Torrance, California, USA). The agar bodies may be coded by shape (e.g. stellate, rectangular, cylindrical), color or embedded e.g. cylindrical color codes or figures (numbers, letters). Thus, the specimens are marked and individualized

independently of the containers at all steps of tissue processing and evaluation. Even at sectioning, staining and microscopic evaluation of the few micrometer thick sections the code provided by the agar bodies can be visualized by naked eye. This holds also true for special staining techniques like immunohistochemistry or fluorescence in situ hybridization.

[0004 ]These basic considerations also apply to plant or animal or inorganic specimens. [0005] Agar is ideally suited for specimen marking because it can be processed (e.g.

paraffinized, cut) like the specimen itself. Agar can be poured or punched in special designs (e.g. stellate, cylindrical), can be colored by commercially available dyes and can contain uni- or multicolored codes e.g. in the form of figures (numbers, letters) or agar cylinders. The cylinders, letters and numbers can be produced in the agar body e.g. by using spacers at pouring the body or by punching the forms into an agar blank. The spaces are then filled with a liquid colored protein-agar-mixture. Native specimens (e.g. breast needle biopsy specimens) may be poured by liquid agar into slots of an agar block containing a coded agar body or sealed together with the coded agar body into a container like the Tissue-Tek® Paraform® Sectionable Cassette System (SAKURA FINETEK USA, Inc., Torrance, California, USA) for the further processing steps.

[0006] The loss of the code provided by the agar body at pretreatment steps of special stainings (e.g. immunohistochemistry, fluorescence in situ hybridization) can be prevented by adding colorized proteins to the agar.

[0007] The novelty of the present invention is the use of agar bodies for an unique marking of specimens by the shape of the agar body, the color or the embedded uni- or multicolored figures (numbers, letters) or agar cylinders. This marking can be read at all steps of sample processing and evaluation e.g. by the naked eye or a scanner with an appropriate software.

[0008] Thus, it is ensured at every processing step that the samples being uniquely marked by the coded agar body are placed into the correspondent containers for further processing. Furthermore, the code may be detected on the sections, providing a correct mounting and evaluation of the sections on the labeled slides.

[0009] By using the present invention a sample mix-up can be prevented from the removal of the sample out of the body till the last processing/evaluation step, e.g. the histological examination under the microscope, by a unique marking of the tissue.

[0010] Other materials that could be used for the identification of tissues are, for example, other biological materials (e.g. animal tissue, cellulose acetate) and cuttable plastics. In order to improve the adhesion of the marking material to the slide in particular in the setting of specific tissue pretreatment procedures e.g. for immunohistochemistry, the agar and the cellulose acetate may be dotted with cells or tissue fragments or other biological materials such as animal tissue or other protein sources can be used. Instead of a chemical dye naturally colored cells (e.g. malignant melanoma cells) can be used for marking.

[0011] These coded agar bodies may be part of a kit which consists of a radiolucent supporting material with slots in which the coded agar bodies and the biological samples may be placed and poured into an agar block by using liquid agar. Thus, the supporting material is used as a mold. The coded agar bodies may contain radiodense substances to make the code visible on a radiogram.

[0012] The prefabricated agar blocks with slots and the incorporated agar body and the cast agar blocks in a kit should have a preferable dimension of 31 x 20 x 4 mm ( length x width x height) and 26 x 20 x 4 mm ( length x width x height) to fit into standard embedding plastic cassettes / standard steel embedding molds or cuttable plastic cassettes (e.g. Sakura Paraform cassettes), respectively. The height of the coded agar body may vary and should correspond to the height of the specimens to be marked. E.g. breast biopsies taken with a 7G and a 14G needle need a body height of about 4 mm and 2 mm, respectively.

[0013] The height of the agar block besides the coded agar body can be small and has only to secure the tight connection between the specimens and the coded agar body. The less height of the agar body facilitates the fixation of the specimens.

[0014] The numbers of the slots per agar block may vary. The dimensions of the slots may be adjusted to the specimens being installed. E.g. a 7G needle biopsy specimen needs more space than a 14G needle biopsy specimen. Moreover, the arrangement of the slots and the coded agar body may vary. The slots and the agar body may be arranged in parallel order or may be arranged perpendicularly. The shape and dimensions of the molds may vary correspondingly to the desired agar block.

[0015] Possible shape variants of the material are displayed in FIGS. 1 (block shaped agar body), 2 (stellate shaped), 3 (cylindrical), 4 (tube), 5 (colored agar body), 6 (agar body with embedded colored number), 7 (agar body with embedded colored letter), 8 (agar body with embedded colored agar cylinders), 9 (agar block with number-coded agar body and with one slot to accommodate the specimen), 10 (agar block with number-coded agar body and with two slots to accommodate the specimens), 11 (agar block with number-coded agar body and with three slots perpendicular to the agar body), 12 (agar block with a number-coded agar body arranged parallel to two slots), 13 (mold to pour prefabricated agar blocks with a coded- agar body and two spacers for producing two slots for the accommodation of the specimens), 14 (mold to pour prefabricated agar blocks with a coded-agar body and two spacers for producing two slots for the accommodation of the specimens), 15 (preformed agar block with incorporated coded agar body and one slot to accommodate the specimen, whereby the height of the coded agar body corresponds to the height of the expected specimen and the height of the rest of the agar block is much smaller), 16 (preformed agar block with incorporated coded agar body and two slots to accommodate the specimens, whereby the height of the coded agar body corresponds to the height of the expected specimen and the height of the rest of the agar block is much smaller), 17 (preformed agar block with incorporated coded agar body and three slots to accommodate the specimens being oriented perpendicular to the agar body, whereby the height of the coded agar body corresponds to the height of the expected specimen and the height of the rest of the agar block is much smaller), 18 (preformed agar block with incorporated coded agar body and two slots to accommodate the specimens being oriented parallel to the coded agar body, whereby the height of the coded agar body corresponds to the height of the expected specimen and the height of the rest of the agar block is much smaller), 19 (Mold for casting an agar block with the coded agar body and the specimens whereby the specimens and the coded agar body are placed in little slots), 20 (Mold for casting an agar block with the coded agar body and the specimens whereby the coded agar body is oriented perpendicular to the length of the agar block), 21 (Mold for casting an agar block with the coded agar body is oriented in parallel to the length of the block), 22 (Paraffin block with a needle biopsy specimen (black irregular structure), paraffinized coded agar body (numeral code) and the processing number (K 12715/15) given by the institute of pathology to mark the specimen containers), 23 (2 μηι thick paraffin section of the paraffin block as shown in Fig. 22 mounted on a glass slide with the processing number at the inscription field of the slide and the needle biopsy specimen with the adjacent unique number of the coded agar body).

Abbreviations used in the figures: w width

h height

1 length

sp spacer