Effects of soft X-ray radiation damage on paraffin-embedded rat tissues supported on ultralene: a chemical perspective

被引:11
作者
Bedolla, Diana E. [1 ]
Mantuano, Andrea [2 ,3 ]
Pickler, Arissa [4 ]
Mota, Carla Lemos [3 ,4 ]
Braz, Delson [4 ]
Salata, Camila [5 ]
Almeida, Carlos Eduardo [3 ]
Birarda, Giovanni [1 ]
Vaccari, Lisa [1 ]
Barroso, Regina Cely [2 ]
Gianoncelli, Alessandra [1 ]
机构
[1] Elettra Sincrotrone Trieste SCpA, SS 14,Km 163,5, I-34149 Trieste, TS, Italy
[2] State Univ Rio de Janeiro UERJ, Phys Inst, Rua Sao Francisco Xavier 524 PJLF Sala 3007F, BR-20550900 Rio De Janeiro, RJ, Brazil
[3] State Univ Rio de Janeiro UERJ, Radiol Sci Lab, Rua Sao Francisco Xavier 524 PHLC Sala 136, BR-20550900 Rio De Janeiro, RJ, Brazil
[4] Univ Fed Rio de Janeiro, COPPE, Av Horacio Macedo 2030,Bloco G Sala 206 CT, BR-21941594 Rio De Janeiro, RJ, Brazil
[5] Comissao Nacl Energia Nucl, Med Phys Dept, Rua Gen Severiano 90, BR-22290901 Rio De Janeiro, RJ, Brazil
关键词
soft X-ray microscopy; radiation damage; Fourier transform infrared (FTIR); paraffin; ultralene; fixation; EMISSION MICROSCOPY; TRANSMISSION; FIXATION; CRYSTALS; BEAMLINE;
D O I
10.1107/S1600577518003235
中图分类号
TH7 [仪器、仪表];
学科分类号
0804 ; 080401 ; 081102 ;
摘要
Radiation damage is an important aspect to be considered when analysing biological samples with X-ray techniques as it can induce chemical and structural changes in the specimens. This work aims to provide new insights into the soft X-ray induced radiation damage of the complete sample, including not only the biological tissue itself but also the substrate and embedding medium, and the tissue fixation procedure. Sample preparation and handling involves an unavoidable interaction with the sample matrix and could play an important role in the radiation-damage mechanism. To understand the influence of sample preparation and handling on radiation damage, the effects of soft X-ray exposure at different doses on ultralene, paraffin and on paraffin-embedded rat tissues were studied using Fourier-transform infrared (FTIR) microspectroscopy and X-ray microscopy. Tissues were preserved with three different commonly used fixatives: formalin, glutaraldehyde and Karnovsky. FTIR results showed that ultralene and paraffin undergo a dose-dependent degradation of their vibrational profiles, consistent with radiation-induced oxidative damage. In addition, formalin fixative has been shown to improve the preservation of the secondary structure of proteins in tissues compared with both glutaraldehyde and Karnovsky fixation. However, conclusive considerations cannot be drawn on the optimal fixation protocol because of the interference introduced by both substrate and embedding medium in the spectral regions specific to tissue lipids, nucleic acids and carbohydrates. Notably, despite the detected alterations affecting the chemical architecture of the sample as a whole, composed of tissue, substrate and embedding medium, the structural morphology of the tissues at the micrometre scale is essentially preserved even at the highest exposure dose.
引用
收藏
页码:848 / 856
页数:9
相关论文
共 29 条
[1]   Infrared spectroscopy of proteins [J].
Barth, Andreas .
BIOCHIMICA ET BIOPHYSICA ACTA-BIOENERGETICS, 2007, 1767 (09) :1073-1101
[2]  
Bozzola J.J, 1999, ELECT MICROSCOPY PRI, V2nd ed.
[3]  
Brown DL., 2016, Atlas of histology of the Juvenile Rat, P173
[4]   THE REACTIONS OF FORMALDEHYDE WITH AMINO ACIDS AND PROTEINS [J].
FRENCH, D ;
EDSALL, JT .
ADVANCES IN PROTEIN CHEMISTRY, 1945, 2 :277-335
[5]   Scanning transmission x-ray microscopy with a configurable detector [J].
Gianoncelli, A. ;
Morrison, G. R. ;
Kaulich, B. ;
Bacescu, D. ;
Kovac, J. .
APPLIED PHYSICS LETTERS, 2006, 89 (25)
[6]   Soft X-Ray Microscopy Radiation Damage On Fixed Cells Investigated With Synchrotron Radiation FTIR Microscopy [J].
Gianoncelli, A. ;
Vaccari, L. ;
Kourousias, G. ;
Cassese, D. ;
Bedolla, D. E. ;
Kenig, S. ;
Storici, P. ;
Lazzarino, M. ;
Kiskinova, M. .
SCIENTIFIC REPORTS, 2015, 5
[7]   Recent developments at the TwinMic beamline at ELETTRA: an 8 SDD detector setup for low energy X-ray Fluorescence [J].
Gianoncelli, A. ;
Kourousias, G. ;
Stolfa, A. ;
Kaulich, B. .
11TH INTERNATIONAL CONFERENCE ON SYNCHROTRON RADIATION INSTRUMENTATION (SRI 2012), 2013, 425
[8]   Simultaneous soft X-ray transmission and emission microscopy [J].
Gianoncelli, A. ;
Kaulich, B. ;
Alberti, R. ;
Klatka, T. ;
Longoni, A. ;
de Marco, A. ;
Marcello, A. ;
Kiskinova, M. .
NUCLEAR INSTRUMENTS & METHODS IN PHYSICS RESEARCH SECTION A-ACCELERATORS SPECTROMETERS DETECTORS AND ASSOCIATED EQUIPMENT, 2009, 608 (01) :195-198
[9]   Current status of the TwinMic beamline at Elettra: asoft X-ray transmission and emission microscopy station [J].
Gianoncelli, Alessandra ;
Kourousias, George ;
Merolle, Lucia ;
Altissimo, Matteo ;
Bianco, Anna .
JOURNAL OF SYNCHROTRON RADIATION, 2016, 23 :1526-1537
[10]   KINETIC-STUDIES OF FORMALDEHYDE BINDING IN TISSUE [J].
HELANDER, KG .
BIOTECHNIC & HISTOCHEMISTRY, 1994, 69 (03) :177-179