title: Evaluation of Histomorphometric Changes in Tissue Architecture in Relation to Alteration in Fixation Protocol - An Invitro Study.
abstract: BACKGROUND
  Preparation of good tissue specimens for microscopy requires complete fixation. No ideal fixative has been found till date, with every fixative showing advantages and disadvantages. Appropriate fixation is required to maintain clear and consistent morphologic features for histologic examination. Pathologists mostly examine formalin fixed tissue sections and are less used to the morphologic changes induced by other fixatives. Underfixed and overfixed tissue in various fixatives can lead to tissue architectural changes which can affect its diagnostic value.
  
  
  OBJECTIVE
  To assess sectioning ability, staining intensity and microscopic details of tissues kept in different fixatives at different time intervals.
  
  
  METHODS
  Fresh tissue specimen i.e., goat tongue was collected and its middle-third portion was used for the study purpose. The tissue was grossed into 10 equal pieces and kept in various fixatives (10% Buffered formalin, Carnoy's solution, Absolute ethyl alcohol, Bouin's fluid) for five different time intervals (6, 12, 18, 24 and 30 hours) and normal tissue processing steps were carried out followed by sectioning and staining. During sectioning, sectioning parameter was assessed. Following sectioning, sections were observed under light microscope and were histologically evaluated for staining and microscopic details. To calculate the sectioning parameter Fisher's exact test was used and to assess parameters for staining and microscopic details Mann-Whitney U test was used.
  
  
  RESULTS
  According to the study, 10% buffered formaldehyde is considered as a superior fixative under all parameters followed by Bouin's fluid, Carnoy's solution and Absolute alcohol.
  
  
  CONCLUSIONS
  In our study, it was concluded that 10% buffered formaldehyde should be continued as a routine fixative however, other fixatives can be used depending upon the non-availability of required fixative or in case of emergencies. Pathologist should be accustomed to histologic and morphologic changes of underfixed and overfixed tissue which can affect its diagnostic value.
authors.affiliation: Senior Lecturer, Department of Oral Pathology and Microbiology, Sarjug Dental College , Darbhanga, Bihar, India .
authors.forename: Pallav
authors.initials: P
authors.lastname: Singhal
authors.affiliation: Professor and Head, Department of Oral Pathology and Microbiology, Kothiwal Dental College , Moradabad, Uttar Pradesh, India .
authors.forename: Narendra Nath
authors.initials: NN
authors.lastname: Singh
authors.affiliation: Professor and Head, Department of Oral Pathology and Microbiology, Babu Banarasi Das College of Dental Sciences , Lucknow, Uttar Pradesh, India .
authors.forename: Gadiputi
authors.initials: G
authors.lastname: Sreedhar
authors.affiliation: Senior Lecturer, Department of Oral Pathology and Microbiology, The Dental College, Regional Institute of Medical Sciences , Imphal, Manipur, India .
authors.forename: Sumita
authors.initials: S
authors.lastname: Banerjee
authors.affiliation: Senior Lecturer, Department of Public Health Dentistry, Surendera Dental College , Sri Ganganagar, Rajasthan, India .
authors.forename: Manu
authors.initials: M
authors.lastname: Batra
authors.affiliation: Postgraduate Student, Department of Pedodontics, Himachal Institute of Dental Science , Paonta Sahib, Himachal Pradesh, India .
authors.forename: Anu
authors.initials: A
authors.lastname: Garg
keywords: Absolute ethyl alcohol; Bouin’s fluid; Buffered formalin; Carnoy’s solution; Incomplete fixation; Over-fixation; Under-fixation