Journal of Physical Science, Vol. 27(3), 85–95, 2016
© Penerbit Universiti Sains Malaysia, 2016
Effect of 460 and 532 nm Laser Light on the Erythrocyte Deformability of Anaemic Blood Samples
Nursakinah Suardi,1* Mohamad Suhaimi Jaafar,1 Mohd Zulkifli Mustafa,2 Abdul Rahim Hussein3 and Zalila Ali4
1School of Physics, Universiti Sains Malaysia, 11800 USM Pulau Pinang, Malaysia
2Department of Neurosciences, School of Medical Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian,
Kota Bharu, Kelantan, Malaysia
3Advanced Medical and Dental Institute, Universiti Sains Malaysia, Bertam Campus, 13200 Kepala Batas, Pulau Pinang, Malaysia
4School of Mathematical Science, Universiti Sains Malaysia, 11800 USM Pulau Pinang, Malaysia
*Corresponding author: nsakinahsuardi@usm.my
Published online: 25 November 2016
To cite this article: Suardi, N. et al. (2016). Effect of 460 and 532 nm laser light on the erythrocyte deformability of anaemic blood samples. J. Phys. Sci., 27(3), 85–95, http://dx.doi.org/10.21315/jps2016.27.3.6
To link to this article: http://dx.doi.org/10.21315/jps2016.27.3.6
ABSTRACT: This study investigated the effect of 460 nm and 532 nm laser irradiation on anaemic blood compared with normal samples at different exposure times. Blood smears were prepared to study the effect of laser irradiation on erythrocyte deformability. Irradiation of normal and anaemic blood samples with 460 and 532 nm laser light significantly changed erythrocyte deformability. The deformability of both the normal and anaemic blood samples increased as the exposure time increased. The analysis also revealed that erythrocyte deformability is greater in anaemic blood than normal blood.
Keywords: Laser, anaemic, erythrocyte, deformability, blood samples
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532 nm Laser L
RESULTS s of both the m to compare
Table 1 pres adiation for hows that la antly change Table 1: E re
s) Control 97.93 ± 0.10 98.20 ±
0.34 86.03 ±
0.10 94.98 ±
0.17 e
l significance: *
rcentage of e s irradiated at nm showed in 90 s time po
ed at 30 s and ood categorie ion.
2 shows the c samples i cyte deforma emic samples amples was u 2. Erythrocyt mal blood sa e normal sa s increased a ability of the
ight
AND DISCU e normal an e these two t ents the effe the normal a aser irradiati ed the erythro Erythrocyte de
Normal samp 460 nm 96.98 ±
0.10 95.00
±0.14 82.00 ±
0.08 92.95 ±
0.06 0.000***
***p<0.01
erythrocyte d t 460 nm is s ncreased def oint. For the d 50 s but sta es, the anaem
percentage rradiated at ability was ob
s irradiated a unaffected af te deformabi amples, with amples, the e
as the expo e normal sam
USSION nd anaemic b
types of bloo ect on erythr and anaemic ion of the n ocyte deform eformability b
ples
532 nm 98.03 ±
0.17 88.00 ±
0.10 81.00 ±
0.08 73.98 ±
0.13 0.000***
deformability shown in Fig formability a
anaemic sam arted to drop mic sample w
of erythrocy t 532 nm.
bserved for t at 460 nm. B fter irradiatio ility increase h the excepti erythrocyte sure time in mples irradiat
blood were i od. The con rocyte deform
samples at 4 normal and mability (p-va
efore and afte
Control 39.03 ± 0.25 91.03 ±
0.13 81.00 ±
0.08 85.00 ±
0.08
y for the no gure 1. The n as the exposu mples, the e p slightly at 7 was affected m
yte deformab After 30 s the normal sa By contrast, t on with the 5 ed as the exp ion of the 7
deformabilit ncreased. Th ted at 460 an
irradiated at trol was the mability befo 460 nm and
anaemic blo alue = 0.000) er laser irradia Anaemic samp
460 nm 41.00 ± 0.09 79.00 ±
0.08 71.00 ±
0.08 78.98 ±
0.17 0.000***
ormal and an normal samp ure time incre
erythrocyte d 70 s and 90 s more by the
bility for the of exposur amples, but i the deformab 532 nm laser, posure time i 0-s exposure ty of the an he highest p nd 532 nm w
88
460 nm and e unirradiated fore and afte
532 nm. The lood sample
).
ation.
mples 532 nm 38.98±0.13
80.97 ± 0.21 81.00 ± 0.08 62.97 ± 0.13
0.000**
naemic blood ples irradiated eased, excep deformability s. Comparing 460 nm lase
e normal and re, only 1%
it was 2% fo bility of both r, as shown in increased fo e time point naemic blood percentage o
was observed
8
d d r e s
3
1 8 3
d d pt y g r
d
% r h n r t.
d f d
Journal of Physical Science, Vol. 27(3), 85–95, 2016 89
at 70 s (4%) and 90 s (21%), respectively. However, for the anaemic samples, the highest deformability was observed at 50 s (12%) and 90 s (22%) when the samples were irradiated at 460 and 532 nm, respectively. At 90 s, irradiation of the blood samples at 532 nm resulted in greater deformability of both the normal and anaemic samples.
Figure 1: Percentage of erythrocyte deformability in normal and anaemic samples irradiated at 460 nm.
Figure 2: Percentage of erythrocyte deformability in normal and anaemic samples irradiated at 532 nm.
0 2 4 6 8 10 12 14
30 50 70 90
Percentage of erythrocyte deformability (%)
Exposure time (s)
Normal samples Anaemic samples
‐5 0 5 10 15 20 25
30 50 70 90
Percentage of erythrocyte deformability (%)
Exposure time (s)
Normal samples Anaemic samples
460 and 532 nm Laser Light 90
The anaemic blood was affected more than the normal blood samples.
Morphologically, both the 460 and 532 nm lasers caused erythrocyte deformability, especially for the anaemic samples. We assumed that the 532 nm laser would affect erythrocyte deformability more than the 460 nm laser, as the greatest deformability percentage was observed after 90 s of irradiation with 532 nm. This effect is because the effective tissue penetration is maximised at 532 nm compared to 460 nm. Haemoglobin, which acts as a chromophore, absorbs more photons at higher absorption bands from the 532 nm laser than the 460 nm laser.9 Thus, the effect is more prominent. Previously, it was shown that laser irradiation improved red blood cell deformability.10,19,20 However, the radiation must be administered at a therapeutic dose and not beyond that.9 The correct dose will stimulate a positive effect, but a higher dose will cause damage to and inhibit the erythrocytes.21
Figure 3 shows images of blood smears from the normal blood samples. Figure 3A is the control unirradiated sample. The erythrocytes in this sample are in good condition. After irradiation for 50 s with the 460 nm laser (Figure 3B) and the 532 nm laser (Figure 3C), deformability of the erythrocytes was observed.
Among the abnormal erythrocytes observed in the smears in Figure 3B and 3C are keratocytes, dacrocytes, boat-shaped cells and echinocytes. Irradiation at 532 nm affected the erythrocytes of the normal samples more than the 460 nm laser.
Figure 4 shows images of blood smears from the anaemic blood samples. Figure 4A is an image of the control unirradiated sample. The erythrocytes from this sample are in poor condition. Abnormal erythrocytes, such as echinocytes and dacrocytes, can already be observed. Irradiation for 50 s with the 460 nm and 532 nm lasers caused more deformability to the erythrocytes than in the control sample, as shown in Figure 4B and 4C, respectively. Among the abnormal erythrocytes observed in the smear in Figure 4B and 4C were schistocytes, acanthocytes, keratocytes, target cells and echinocytes.
Based on all the blood smears prepared, most of the abnormal cells observed were echinocytes, which are crenated compared to the other abnormal red blood cell variants. The echinocytes can become spherocytes as they lose membrane vesicles, which leads to a greater loss of surface area and volume and finally leads to haemolysis.22–24 Therefore, a suitable laser irradiation parameter must be carefully chosen after weighing the benefits and the risks.
The red blood cell lifespan depends on an adequate oxidative stress response. Red blood cells are made of protein, and denaturation can occur when the local heat is high, which causes shear stress to the red blood cell membrane and thus changes
Journal of Physical Science, Vol. 27(3), 85–95, 2016 91
the shape of the red blood cell. Echinocytes also form because of decreased ATP generation, resulting in the loss of water and potassium from the red blood cells.
Red blood cells have deformable structures that allow them to recover their initial shape after passing through very small capillaries, which is an important and essential feature for their blood flow properties.14,18,25,26 However, if the alteration and deformation of the red blood cell is severe, it may affect the ability of the cell to function properly. Thus, maintaining the percentage of deformability of red blood cells after laser irradiation is crucial.
Figure 3: Images of blood smears from normal blood samples of A) control unirradiated sample; B) irradiated at 460 nm; and C) irradiated at 532 nm.
460 and 5
Figure 4
4.
Red blo whole b lasers si normal a analysis than th paramet
532 nm Laser L
: Images of b sample; B)
CONCLUS ood cells are body. Irradiat ignificantly and anaemic s also reveale
at of norm ter must be c
ight
blood smears irradiated at 4
SION important ce ting normal a changes the c blood samp ed that the d mal blood sa
carefully cho
from normal 460 nm; and C
ells for hum and anaemic eir deformab ples increased deformability
amples. The sen after wei
blood sample C) irradiated a
mans that regu c blood samp bility. The d
d as the expo y of anaemic erefore, a s ighing the be
es of A) contro at 532 nm.
ulate the fun ples with 460 eformability osure time in blood samp uitable lase enefits and th
92
ol unirradiated
nctions of the 0 and 532 nm y of both the ncreased. The ples is greate
er irradiation he risks.
2
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Journal of
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The aut providin Haemat Neurosc support Ethics C Reg. No
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ACKNOWL thors wish t ng the resear
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of this rese Committee, o: 00004494)
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