Gorgonian and scleractinian coral t

Gorgonian and scleractinian coral tissue sample
processing
Following decalcification, gorgonian and scleractinian coral
tissue samples were washed in filtered seawater twice (10
min each). Then, tissues were placed in the microwave for
dehydration in a sequence of 95% ethanol (Fisher Scientific,
Waltham, MA, 95% ethanol: 100% isopropanol (2-propanol
hereafter) [Fisher Scientific, Waltham, MA], and cleared in 100%
2-propanol. Each step lasted 4 min and temperature was set
to 40oC. Next, tissue samples were kept in the microwave for
infiltration in a 1:1 solution of melted paraffin and 2-propanol
under vacuum for 10 min at 60oC (Table 1). A second round of
infiltration was performed with 100% melted paraffin under
vacuum for 30 min at 60oC. During the infiltration process, a
sheet of PolarHeat™ (Energy Beam Sciences, CT) was used to
convert microwave energy to heat, resulting in an increase in
diffusivity of the paraffin through the tissue [14].
Sectioning and hematoxylin and eosin (H&E) staining
Thin sections (4-5μm) from gorgonian and scleractinian
coral tissue were obtained using a rotary microtome (model
820; American Optical, Buffalo, NY). Sections were affixed to
subbed-glass slides (chrome alum-gelatin coating solution:
50.0 g gelatin (Mallinckrodt, St. Louis, MO) and 5.0 g chromium
potassium sulfate (Mallinckrodt, St. Louis, MO) in 1 L distilled
deionized water.
Sections were deparaffinized with Histo-Clear twice, (30 s
each, National Diagnostics, Charlotte, NC), re-hydrated through
a descending series of ethanols (95%-70% ethanol each for 30
s), washed in distilled deionized water (2X each 30 s), stained
in Harris hematoxylin (C.I. #75290, Sigma Aldrich, St. Louis,
MO) solution for 1 min, washed in running tap water for 1
min, differentiated in Scott solution (10g sodium bicarbonate;
2g magnesium sulfate in 1 L distilled deionized water; [15])
for 30 s, and washed in running tap water again for 1 min.
Bluing was done using 0.2% ammonia water for 30 s to 1 min.

Hasil (Bahasa Indonesia) 1:

Sampel jaringan karang gorgonian dan scleractinianpengolahanBerikut karang dekalsifikasi, gorgonian dan scleractiniansampel jaringan itu dicuci di disaring air dua kali (10menit masing-masing). Kemudian, Jaringan ditempatkan dalam microwave selamadehidrasi dalam urutan yang 95% etanol (Fisher ilmiah,Waltham, MA, 95% etanol: 100% isopropanol (2-propanolakhirat) [Fisher Scientific, Waltham, MA], dan dihapus dalam 100%2-propanol. Setiap langkah yang berlangsung selama 4 menit dan suhu ditetapkanuntuk 40oC. Selanjutnya, sampel jaringan dipelihara dalam microwave selamainfiltrasi dalam larutan 1:1 parafin meleleh dan 2-propanoldi bawah vakum untuk 10 min di 60oC (Tabel 1). Putaran keduainfiltrasi dilakukan dengan parafin 100% meleleh di bawahvakum selama 30 menit di 60oC. Selama proses infiltrasi,lembar PolarHeat ™ (energi sinar ilmu, CT) digunakan untukmengubah microwave energi panas, mengakibatkan peningkatandiffusivity parafin melalui jaringan [14].-Sectioning dan hematoxylin dan eosin (H & E) nodaBagian tipis (4-5μm) dari gorgonian dan scleractiniankarang jaringan diperoleh dengan menggunakan rotary microtome (model820; Amerika optik, Buffalo, NY). Bagian yang ditempelkankaca-subbed slide (tawas chrome-gelatin lapisan solusi:gelatin 50,0 g (Mallinckrodt, St Louis, MO) dan kromium 5.0 gKalium sulfat (Mallinckrodt, St Louis, MO) dalam 1 L sulingdeionized air.Bagian deparaffinized dengan jelas Histo dua kali, (30 smasing-masing, diagnostik nasional, Charlotte, NC), kembali terhidrasi melaluiserangkaian menurun ethanols (95% - 70% etanol masing-masing sebesar 30s), dicuci dalam air suling deionized (2 X 30 setiap s), bernodadi Harris hematoxylin (CI #75290, Sigma Aldrich, St Louis,MO) solusi untuk 1 min, dicuci dalam air keran untuk 1Min, dibedakan dalam larutan Scott (10g natrium bikarbonat;2g magnesium sulfat di 1 L suling deionized air; [15])untuk 30 s, dan dicuci dalam air keran berjalan lagi untuk 1 menit.Bluing dilakukan menggunakan 0,2% amonia air selama 30 s untuk 1 min.