Humans ; Floods ; Paraffin ; Paraffin Embedding

BACKGROUND: Inevitable loss of diagnostic material should be minimized during cell block preparation. We introduce a modified agarose cell block technique that enables the synthesis of compact cell blocks by using the entirety of a cell pellet without the loss of diagnostic material during cell block preparations. The feasibility of this technique is illustrated by high-throughput immunocytochemistry using high-density cell block microarray (CMA). METHODS: The cell pellets of Sure- Path residues were pre-embedded in ultra-low gelling temperature agarose gel and re-embedded in standard agarose gel. They were fixed, processed, and embedded in paraffin using the same method as tissue sample processing. The resulting agarose cell blocks were trimmed and represented on a CMA for high-throughput analysis using immunocytochemical staining. RESULTS: The SurePath residues were effectively and entirely incorporated into compact agarose cell buttons and embedded in paraffin. Sections of the agarose cell blocks revealed cellularities that correlated well with corresponding SurePath smears and had immunocytochemical features that were sufficient for diagnosis of difficult cases. CONCLUSIONS: This agarose-based compact cell block technique enables preparation of high-quality cell blocks by using up the residual SurePath samples without loss of diagnostic material during cell block preparation.
Biopsy, Fine-Needle ; Diagnosis ; Immunohistochemistry ; Paraffin ; Paraffin Embedding ; Sepharose*

Disinfectants ; Fixatives ; toxicity ; Formaldehyde ; toxicity ; Paraffin Embedding ; Tissue Fixation

Paraffin Embedding ; methods ; Tissue Array Analysis ; instrumentation ; methods

Humans ; Paraffin Embedding ; methods ; Periodontium ; pathology ; Tooth ; pathology

The partial splenectomy was performed on the basis of the arterial distribution in order to reduce complications. The spleen consists of the various histologic components that have different functions. But we can not find the reports whether the distribution of red pulp, white pulp and trabecula is uniform or not according to the regions of the spleen. We used 15 spleens from Korean[male 8, female 7 and age 17-74]. The volume was measured with the mass cylinder. The 1cm3 blocks were selected in 5 different regions. Whole splenic slices by celloidin embedding were made from two spleens. The point counting method with eyepeice reticule was used for the measurement of the distribution of red pulp, white pulp and trabecula. The results were as follows : 1. The volume of the spleen was ranged 45-158ml and the individual difference was marked. The male spleen was larger than female`s [p<0.05]. 2. The ratio of red pulp, white pulp and trabecula of the paraffin embedding preparations was 83.2, 10.9, and 5.9% respectively. There was no significant difference in the distribution of the histologic components among five regions[upper, hilum, middle, outer, and lower]. 3. The distribution of red and white pulps in whole splenic slices was not uniformity. The ratio of red pulp, white pulp and trabecula in whole splenic slices was 80.1, 13.4 and 6.5% respectively. These results showed no significant difference with those of five regions of the spleen.
Collodion ; Female ; Humans ; Individuality ; Male ; Paraffin Embedding ; Spleen* ; Splenectomy

Biopsy ; Cryopreservation ; Frozen Sections ; Humans ; Kidney ; pathology ; Kidney Diseases ; pathology ; Paraffin Embedding ; methods

Formaldehyde ; Humans ; Microdissection ; Paraffin Embedding ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Tissue Fixation

OBJECTIVETo explore the feasibility of detecting lymphoma with the application of BIOMED-2 standardized immunoglobulin/T cell receptor (IG/TCR) gene rearrangement system in formalin fixed paraffin-embedded (FFPE) tissue samples, and to discuss the relationship between the longest amplification fragment of extracted DNA and positive detection rate of different IGH V-J primers.

METHODSDNA was extracted from 50 cases of FFPE tissue samples. Multiplex-PCR amplifications were performed and then the IG/TCR gene rearrangements were analyzed using BIOMED-2 standardized clonality analysis system.

RESULTS(1)When the DNA concentration was diluted to 50-100 ng/μl from 100-500 ng/μl, the proportion of the longest amplification fragment (300-400 bp) of DNA was improved from 10.0% to 90.0% in 30 cases of diffuse large B cell lymphoma (DLBCL) wax roll samples (P<0.01). The positive rate of IGH+IGK was increased from 46.7% to 83.3%, the difference was statistically significant (P=0.006). The lengths of the longest amplification fragments of DNA were all longer than 300 bp in the paraffin section samples of DLBCL. The positive rate of IGH+IGK of these samples was 96.7%. The difference of the positive rate of IGH+IGK between the wax roll samples and the paraffin section samples had no statistical significance (P=0.195). (2)When the concentration of DNA was high, most of the longest amplification fragments of extracted DNA were 100 bp or 200 bp, and the detection rate of short fragment IGH FR3 was more stable than that of long fragment IGH FR1. (3)The clonality analysis of TCRG+TCRB in all 13 cases of peripheral T cell lymphoma samples showed positive results, while no positive IG/TCR clones were found in 7 cases of reactive lymphoid tissue hyperplasia in control group.

CONCLUSIONDilution of DNA is the only method to improve not only the proportion of longest fragment amplification but also the detection rate of clonality. The detection rate of IGH FR3 would not be affected by the concentration of DNA. The application of BIOMED-2 standardized IG/TCR gene rearrangement system in FFPE tissue samples plays an important role in the lymphoma diagnosis.

Biopsy ; Histological Techniques ; instrumentation ; methods ; Humans ; Indicators and Reagents ; Paraffin Embedding ; Ultrasonics