The employed method was indirect im

The employed method was indirect immunohistochemical
staining that involves an unlabeled primary
antibody specific to the target antigen. An enzymatically
labeled secondary antibody reacted with the primary antibody,
and the reaction was visualized with a chromogenic
substrate.
The study was performed on paraffin-embedded sections
from skin biopsies collected from patients with acne
vulgaris, the patient diagnosed with AF included.
After washing with distilled water, the sections were
counterstained with Hematoxylin for two minutes at
room temperature, followed by washing with distilled
water, 100% ethanol and benzene/xylene.
Sections were investigated with a Nikon microscope
and the presence of specific markers was highlighted by
the appearance of a brown precipitate in the membrane
(CD3 and CD20) or cytoplasm (cytokeratins, CD68,
CD34 II) corresponding to the identified antigen. Specific
marking of the epithelial cells by anti-cytokeratin antibody
was evaluated qualitatively positive versus negative, for
different types of lesions or epithelial structures present
in the skin biopsies. We established three degrees of
cytokeratin immunostaining intensity: + (weak intensity),
++ (medium intensity), and +++ (high intensity).
The inflammatory infiltrate was assessed for each
staining by counting the marked cells and expressing the
results as a percentage of total cells present in a microscopic
field.
Other sections were subjected to the same protocol,
except for the primary antibody incubation step, which
were replaced with antibodies of the same isotype, but
with irrelevant specificity, serving as negative controls.
In the studied acne cases, AF included, cytokeratin
staining was present in the epidermal keratinocytes, cells
of pilosebaceous follicle, sebaceous and apocrine sweat
glands (Figure 5).

Results (Indonesian) 1:

Metode yang digunakan adalah langsung Imunohistokimiapewarnaan yang melibatkan dasar unlabeledantibodi spesifik untuk target antigen. Enzimatik padalabel sekunder antibodi bereaksi dengan antibodi utama,dan reaksi divisualisasikan dengan chromogenicsubstrat.Penelitian dilakukan pada bagian parafin-tertanamdari biopsi kulit yang dikumpulkan dari penderita jerawatvulgaris, pasien didiagnosis dengan AF disertakan.Setelah mencuci dengan air suling, Bagian yangcounterstained dengan Hematoxylin selama dua menit padasuhu kamar, diikuti dengan mencuci dengan sulingair, 100% etanol dan benzena xilena.Bagian diselidiki dengan mikroskop Nikondan adanya tanda-tanda spesifik disoroti olehmunculnya percepatan cokelat dalam membran(CD3 dan CD20) atau sitoplasma (cytokeratins, CD68,CD34 II) sesuai dengan antigen diidentifikasi. Spesifikmenandai dari sel-sel epitel oleh antibodi anti-cytokeratindievaluasi secara kualitatif positif versus negatif, untukJenis lesi atau epitel struktur hadirpada biopsi kulit. Kami mendirikan tiga derajat daricytokeratin immunostaining intensitas: + (lemah intensitas),++ (intensitas sedang), dan +++ (intensitas tinggi).Menyusup inflamasi dinilai untuk masing-masingpewarnaan oleh menghitung sel ditandai dan mengungkapkanhasil sebagai persentase dari total sel hadir di mikroskopisbidang.Bagian lain adalah tertakluk kepada protokol yang sama,except for the primary antibody incubation step, whichwere replaced with antibodies of the same isotype, butwith irrelevant specificity, serving as negative controls.In the studied acne cases, AF included, cytokeratinstaining was present in the epidermal keratinocytes, cellsof pilosebaceous follicle, sebaceous and apocrine sweatglands (Figure 5).