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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

Construction of a cell block microarray (CMA) for high-throughput immunocytochemistry of cell blocks. A 3-mm tissue punch is used to extract cores of agarose cell buttons from the agarose cell blocks and implant them in a ready-to-use homemade recipient agarose paraffin block (A, B). The CMA is completely melted on a heat plate to facilitate complete integration of cell block cores into the recipient agarose paraffin block (C, D). Finally, the CMA is re-embedded in paraffin and trimmed to expose the cell block cores (E, F).
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f3-kjpathol-48-5-351: Construction of a cell block microarray (CMA) for high-throughput immunocytochemistry of cell blocks. A 3-mm tissue punch is used to extract cores of agarose cell buttons from the agarose cell blocks and implant them in a ready-to-use homemade recipient agarose paraffin block (A, B). The CMA is completely melted on a heat plate to facilitate complete integration of cell block cores into the recipient agarose paraffin block (C, D). Finally, the CMA is re-embedded in paraffin and trimmed to expose the cell block cores (E, F).

Mentions: To save time and resources in examining the applicability of the agarose cell blocks to adjuvant immunocytochemistry, the cores of paraffin-embedded agarose cell buttons in the agarose cell blocks were represented in a CMA[16]. In order to incorporate the entire cases into a single CMA, we used a self-made manual microarray kit and a homemade recipient block as described previously (Fig. 3)[17,18]. Tissue cores from redundant FFPET were also arrayed in the same CMA to be used as macroscopic orientation markers of the CMA sections. Serial sections were cut from the CMA and routinely processed for H&E and immunostaining. An automated slide stainer (Ventana BenchMark XT, Ventana Medical Systems Inc., Tucson, AZ, USA) and the Ventana OptiView DAB detection kit were used to analyze expression of markers (Table 1) using the same antibodies and protocols standardized for immunostaining of FFPET sections. A digital slide scanner (VM600, Motic, Xiamen, China) was used to acquire the entire image of the H&E and immunostained CMA sections.


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)

Construction of a cell block microarray (CMA) for high-throughput immunocytochemistry of cell blocks. A 3-mm tissue punch is used to extract cores of agarose cell buttons from the agarose cell blocks and implant them in a ready-to-use homemade recipient agarose paraffin block (A, B). The CMA is completely melted on a heat plate to facilitate complete integration of cell block cores into the recipient agarose paraffin block (C, D). Finally, the CMA is re-embedded in paraffin and trimmed to expose the cell block cores (E, F).
© Copyright Policy
Related In: Results  -  Collection

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

f3-kjpathol-48-5-351: Construction of a cell block microarray (CMA) for high-throughput immunocytochemistry of cell blocks. A 3-mm tissue punch is used to extract cores of agarose cell buttons from the agarose cell blocks and implant them in a ready-to-use homemade recipient agarose paraffin block (A, B). The CMA is completely melted on a heat plate to facilitate complete integration of cell block cores into the recipient agarose paraffin block (C, D). Finally, the CMA is re-embedded in paraffin and trimmed to expose the cell block cores (E, F).
Mentions: To save time and resources in examining the applicability of the agarose cell blocks to adjuvant immunocytochemistry, the cores of paraffin-embedded agarose cell buttons in the agarose cell blocks were represented in a CMA[16]. In order to incorporate the entire cases into a single CMA, we used a self-made manual microarray kit and a homemade recipient block as described previously (Fig. 3)[17,18]. Tissue cores from redundant FFPET were also arrayed in the same CMA to be used as macroscopic orientation markers of the CMA sections. Serial sections were cut from the CMA and routinely processed for H&E and immunostaining. An automated slide stainer (Ventana BenchMark XT, Ventana Medical Systems Inc., Tucson, AZ, USA) and the Ventana OptiView DAB detection kit were used to analyze expression of markers (Table 1) using the same antibodies and protocols standardized for immunostaining of FFPET sections. A digital slide scanner (VM600, Motic, Xiamen, China) was used to acquire the entire image of the H&E and immunostained CMA sections.

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