[2115] Objective Histologic Stain Quality and Variability Analysis through Digital Imaging: The Effect of Staining Automation

Matthew J Swadley, Marvin L Jones, Alton B Farris. Emory University, Atlanta, GA

Background: Histologic staining variability often poses difficulty in pathologic diagnosis and can be a source of frustration for practicing pathologists. However, assessment is often based upon subjective eyeball evaluation of glass slides. Digital image analysis provides the ability to objectively quantify such evaluations, and allow for regimented quality control over this portion of the hospital laboratory.
Design: Twenty-four daily Masson's Trichrome control slides (12 hand-stained, 12 automated) from available nearest consecutive days prior to and after the implementation of automated staining were digitally scanned with an Aperio Scanscope XT. Two 10X screenshots of representative vasculature were captured from each whole slide file in Aperio Imagescope. Using the open source GNU Image Manipulation Program (GIMP), the optical color components of hue, saturation, and intensity value (HSV) were systematically recorded for 10 points of perivascular fibrous tissue of each vessel. Combined HSV measurements were averaged for each day and analyzed for mean deviation and day-to-day variability.
Results: Automation resulted in statistically significant differences in both stain appearance and day-to-day variability. Mean perivascular stain hue and saturation differed significantly (p = 0.002 and 0.0006, respectively), while value showed no significant change.

Mean HSV measurements
Hue (95% CI)250.4 (237.7-263.1)222.7 (218.9-226.6)
Saturation (95% CI)43.5 (36.9-50.1)65.1 (57.9-72.5)
Value (95% CI)64.3 (60.0-68.6)63.7 (60.2-67.2)
*Hue, saturation, and value are unitless measurements

Additionally, day-to-day hue variability was significantly decreased using an automated stainer (Pre- and post-automation hue standard deviations 22.4 and 6.8, respectively).

Conclusions: Initiation of automated staining yielded significant changes in hue and saturation and decreased day-to-day staining variability. Digital HSV analysis can be a viable method of objectively assessing stain quality and variability within the histology laboratory. Using acceptable hue and saturation error levels, histologic stain quality can be quantitatively assessed in similar fashion to analytical tests within the clinical laboratory.
Category: Quality Assurance

Monday, March 19, 2012 1:00 PM

Poster Session II # 283, Monday Afternoon


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