A new and standardized method to sample and analyse vitreous samples by the Cellient automated cell block system

Acta Ophthalmol. 2014 Aug;92(5):e388-92. doi: 10.1111/aos.12360. Epub 2014 Feb 7.

Abstract

Purpose: In this prospective study, a universal protocol for sampling and analysing vitreous material was investigated. Vitreous biopsies are difficult to handle because of the paucity of cells and the gelatinous structure of the vitreous. Histopathological analysis of the vitreous is useful in difficult uveitis cases to differentiate uveitis from lymphoma or infection and to define the type of cellular reaction.

Methods: Hundred consecutive vitreous samples were analysed with the Cellient tissue processor (Hologic). This machine is a fully automated processor starting from a specified container with PreservCyt (fixative fluid) with cells to paraffin. Cytology was compared with fixatives Cytolyt (contains a mucolyticum) and PreservCyt. Routine histochemical and immunostainings were evaluated.

Results: In 92% of the cases, sufficient material was found for diagnosis. In 14%, a Cytolyt wash was necessary to prevent clotting of the tubes in the Cellient due to the viscosity of the sample. In 23%, the diagnosis was an acute inflammation (presence of granulocytes); in 33%, chronic active inflammation (presence of T lymphocytes); in 33%, low-grade inflammation (presence of CD68 cells, without T lymphocytes); and in 3%, a malignant process.

Conclusion: A standardized protocol for sampling and handling vitreous biopsies, fixing in PreservCyt and processing by the Cellient gives a satisfactory result in morphology, number of cells and possibility of immuno-histochemical stainings. The diagnosis can be established or confirmed in more than 90% of cases.

Keywords: Cellient®; automated system; cell block; vitreous biopsies.

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Antigens, CD / metabolism
  • Antigens, Differentiation, Myelomonocytic / metabolism
  • Biopsy
  • Female
  • Granulocytes / pathology
  • Histocytological Preparation Techniques / methods*
  • Humans
  • Ki-67 Antigen / metabolism
  • Lymphoma, B-Cell / diagnosis*
  • Lymphoma, B-Cell / metabolism
  • Male
  • Melanoma / diagnosis*
  • Melanoma / metabolism
  • Paraffin Embedding
  • Prospective Studies
  • Specimen Handling
  • T-Lymphocytes / pathology
  • Uveitis / diagnosis*
  • Uveitis / metabolism
  • Vitrectomy
  • Vitreous Body / pathology*

Substances

  • Antigens, CD
  • Antigens, Differentiation, Myelomonocytic
  • CD68 antigen, human
  • Ki-67 Antigen