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Preparation of compact agarose cell blocks from the residues of liquid-based cytology samples.

Choi SJ, Choi YI, Kim L, Park IS, Han JY, Kim JM, Chu YC - Korean J Pathol (2014)

Bottom Line: The resulting agarose cell blocks were trimmed and represented on a CMA for high-throughput analysis using immunocytochemical staining.The SurePath residues were effectively and entirely incorporated into compact agarose cell buttons and embedded in paraffin.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Inha University Hospital, Inha University School of Medicine, Incheon, Korea.

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus

SurePath smears and corresponding agarose cell blocks of serous effusions. The cellularities in SurePath smears of serous effusions with reactive mesothelial proliferation (A), malignant mesothelioma (B), and metastatic adenocarcinoma (C) correlate well with those of corresponding agarose cell block sections (D-F), respectively.
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f5-kjpathol-48-5-351: SurePath smears and corresponding agarose cell blocks of serous effusions. The cellularities in SurePath smears of serous effusions with reactive mesothelial proliferation (A), malignant mesothelioma (B), and metastatic adenocarcinoma (C) correlate well with those of corresponding agarose cell block sections (D-F), respectively.

Mentions: By using ULGT agarose solution as a resuspending medium, the residual diagnostic materials in the residual SurePath sample was easily and entirely incorporated into a compact agarose cell button which measured 3 to 5 mm in diameter. By using the cap of an Eppendorf reaction tube as an embedding mold and the standard agarose as a pre-embedding medium, the compact agarose cell button was easily integrated at the base of an agarose gel disk, which was easy to manipulate for further procedures for tissue processing and paraffin embedding. Following the routine 12- to 13-hour tissue processing, all of the agarose gel disks were well impregnated with paraffin and retained their original size and shape. With the aid of FFPET embedded in parallel or a marking dye applied in advance to the agarose cell buttons, it was not difficult for the histotechnologists to determine the optimal cutting level of the cell blocks. Although a thin layer of nearly transparent agarose gel was present in the H&E-stained sections, it did not obscure the cytologic and architectural features because it was retained outside the cells or tissue fragments. The cytomorphologic features of the cell block sections were supportive for the original diagnosis of the corresponding SurePath smears (Figs. 4, 5).


Preparation of compact agarose cell blocks from the residues of liquid-based cytology samples.

Choi SJ, Choi YI, Kim L, Park IS, Han JY, Kim JM, Chu YC - Korean J Pathol (2014)

SurePath smears and corresponding agarose cell blocks of serous effusions. The cellularities in SurePath smears of serous effusions with reactive mesothelial proliferation (A), malignant mesothelioma (B), and metastatic adenocarcinoma (C) correlate well with those of corresponding agarose cell block sections (D-F), respectively.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4215960&req=5

f5-kjpathol-48-5-351: SurePath smears and corresponding agarose cell blocks of serous effusions. The cellularities in SurePath smears of serous effusions with reactive mesothelial proliferation (A), malignant mesothelioma (B), and metastatic adenocarcinoma (C) correlate well with those of corresponding agarose cell block sections (D-F), respectively.
Mentions: By using ULGT agarose solution as a resuspending medium, the residual diagnostic materials in the residual SurePath sample was easily and entirely incorporated into a compact agarose cell button which measured 3 to 5 mm in diameter. By using the cap of an Eppendorf reaction tube as an embedding mold and the standard agarose as a pre-embedding medium, the compact agarose cell button was easily integrated at the base of an agarose gel disk, which was easy to manipulate for further procedures for tissue processing and paraffin embedding. Following the routine 12- to 13-hour tissue processing, all of the agarose gel disks were well impregnated with paraffin and retained their original size and shape. With the aid of FFPET embedded in parallel or a marking dye applied in advance to the agarose cell buttons, it was not difficult for the histotechnologists to determine the optimal cutting level of the cell blocks. Although a thin layer of nearly transparent agarose gel was present in the H&E-stained sections, it did not obscure the cytologic and architectural features because it was retained outside the cells or tissue fragments. The cytomorphologic features of the cell block sections were supportive for the original diagnosis of the corresponding SurePath smears (Figs. 4, 5).

Bottom Line: The resulting agarose cell blocks were trimmed and represented on a CMA for high-throughput analysis using immunocytochemical staining.The SurePath residues were effectively and entirely incorporated into compact agarose cell buttons and embedded in paraffin.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Inha University Hospital, Inha University School of Medicine, Incheon, Korea.

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus