Blue light‐emitting diode technology‐operated microscopy is preferable to both short arc mercury lamp‐operated microscopy and laser scanning confocal microscopy for direct immunofluorescence images evaluation in routinely diagnosing subepidermal autoimmune blistering diseases

Abstract

Direct immunofluorescence (DIF) microscopy still is the gold standard in diagnosing and differentiating subepidermal autoimmune blistering diseases (SABDs) from other bullous diseases. New optical systems were developed that aim to facilitate DIF images evaluation. The aim of the study was to evaluate the usefulness of three fluorescence microscopy systems with different light source for routine diagnostics of SABDs with DIF. In total, perilesional tissue samples for DIF from 34 SABD patients were examined under the three commercial microscopy systems (short arc mercury lamp‐operated microscopy—MLM, blue light‐emitting diode technology‐operated microscopy—bLED and laser scanning confocal microscopy—LSCM) for the detection and pattern analysis of IgA, IgG, IgG1, IgG4, C3 immunoreactants along the dermal–epidermal junction (DEJ) by three independent observers simultaneously. MLM, bLED, and LSCM provided comparable quality of disease‐characterizing immunoreactants imaging (number of immunoreactant types detected and patterns of their deposition along DEJ were the same) but screening of slides was quicker using bLED than both LSCM and MLM, as statistical analysis indicated. It is concluded that bLED is an efficient and preferable system for routinely diagnosing SABDs cost‐effectively. Research Highlights New optical systems were developed that aim to facilitate direct immunofluorescence evaluation. Here, we evaluate the usefulness of three fluorescence microscopy systems with different light source for routine diagnostics concluding that that bLED is an efficient and preferable system.

An update on direct immunofluorescence for diagnosing dermatitis herpetiformis

“…DIF can be visualized with short arc mercury lamp-operated microscopy, blue light-emitting diode technologyoperated microscopy and laser scanning confocal microscopy [5] (Figure 1). Conceptually, a super/high resolution microscopy technique known under the acronym STED (stimulated emission depletion) [6], if adapted for the routine laboratory use, should also be useful for evaluation DIF images, especially as nowadays it can be fixed to the microscope of any type utilizing just the shoebox size equipment.…”

Conceptualization and validation of an innovative direct immunofluorescence technique utilizing fluorescein conjugate against IgG + IgG4 for routinely diagnosing autoimmune bullous dermatoses

“…In order to limit subjectivity, the assessment of DIF specimens for unequivocal identification of the types of immunoreactants present was performed by two independent evaluators, using two different fluorescence microscopic systems: blue light-emitting diode technology-operated microscopy (EuroStar III Plus microscope, Euroimmun, Germany) (concluded to be preferable for routine laboratory diagnostics of ABDs) and short arc mercury lamp-operated microscopy (BX40, Olympus, Japan) at identical objective magnifications (20×, 40×) [ 25 ].…”

A Comparative Analysis of CD32A and CD16A Polymorphisms in Relation to Autoimmune Responses in Pemphigus Diseases and Subepithelial Autoimmune Blistering Disorders

“…The slides were then coverslipped and examined. Skin samples were examined by up to three independent observers using up to three methods, including blue light-emitting diode technology-operated microscopy (EuroStar III Plus microscope, Euroimmun, Germany), short arc mercury lamp-operated microscopy (BX40, Olympus, Japan) and laser-scanning confocal microscopy (the ZEISS LSM510 system with Axiovert 200M laser-scanning confocal microscope, Carl Zeiss Jena GmbH, Germany) [24].…”