1.5 Nanoparticles
1.5.1 Fe3O4 is nature magnet
1.5.1 Fe3O4 is nature magnet
It likes nature little magnet in the body. Most materials found in organisms are generally thought of as being nonmagnetic- for example, C.elegans[13],either diamagnetic (repelled weakly from a magnetic field, as is water and almost any fatty substance) or paramagnetic (weakly attracted to a magnetic field, as is deoxyhemoglobin in blood cells [21].
1.5.2 TiO2, Fe2O3 and Fe3O4 toxicity
Fe3O4, Fe2O3, and TiO2 had no measurable effect on the cells toxicity. Toxicity of iron oxides and metabolites of benzo apyrene alone or in combination in cells culture and identification by laser microprobe mass spectrometry.[22] These results suggest that Fe2O3 and Fe3O4 alone are not very toxic but the association of one of these compounds with BaP increases the toxicity of the latter. On the other hand, TOF-LMMS seems to show a metabolization of iron oxide into reduced form. But, it is necessary to raise the ambiguity about the iron which is always in the cells. \TiO2 had no measurable effect on the cells until the concentrations reached greater than 200 microg/mL [23].
LDH leakage significantly increased in the cells exposed to ZnO (50 to 100 microg/mL), while other nanoparticles tested displayed LDH leakage only at higher doses (>200 microg/mL).
1.6 Magnetic field and Gene expression
Exposure to magnetic field (5 mT at 60 Hz) does not affect cell growth and c-myc gene expression designed and manufactured equipments for long-term and low-density (0 to 9 mT) exposures of cultured cells to extremely low frequency magnetic fields (ELF-MF), and examined the effects of ELF-MF on cell growth and c-myc mRNA expression in Chinese hamster ovary (CHO) cells [16].
The wide-spread induction of stress-related genes and transcription factors, and a depression of genes associated with cell wall metabolism, are prominent examples. Magnetic Field-Responsive Sequences in hsp70. [18] HSP70 gene expression is induced by a wide range of environmental stimuli, including 60-Hz electromagnetic fields. In an earlier report we showed that the induction of HSP70 gene expression by magnetic fields is effected at the level of transcription and is mediated through c-myc protein binding at two nCTCTn sequences at 2230 and 2160. In the human HSP70 promoter. report on the identification of a third c-myc binding site (between 2158 and 2162) that is an important regulator of magnetic field-induced HSP70 expression.We also show that the heat shock element (HSE), lying between 2180 and 2203, is required for induction of HSP70 gene expression by magnetic fields[20]. High magnetic field induced changes of gene expression in
Arabidopsis[19) Field strengths in excess of about 15 Tesla induce expression of the Adh/GUS transgene in the roots and leaves. From the microarray analyses that surveyed 8000 genes, 114 genes were differentially expressed to a degree greater than 2.5 fold over the control. These results werequantitatively corroborated by qRT-PCR examination of 4 of the 114 genes [19].
1.7 Apoptosis and gene
Apoptosis is mediated by a family of cysteine-aspases (caspases), which are expressed as inactive zymogens and are prototypically processed to an active state following an apoptotic stimulus. Many important human diseases are caused by abnormalities in the control of cellular apoptosis (programmed cell death). These abnormalities can result in either a pathological increase in cell number (e.g. cancer) or a damaging loss of cells (e.g. degenerative diseases).
1.7.1 Human apoptosis related gene and apoptosis pathway
The intrinsic pathway (Fig 2) requires disruption of the mitochondrial membrane and the release of mitochondrial proteins including Smac/DIABLO, HtRA2, and cytochrome c. Cytochrome c functions with Apaf-1 to induce activation of caspase-9, thereby initiating the apoptotic caspase cascade, while Smac/DIABLO and HtrA2 bind to and antagonize IAPs. Mitochondrial membrane permeabilization is regulated by the opposing actions of pro- and antiapoptotic Bcl-2 family members. Multidomain proapoptotic Bcl-2 proteins (e.g., Bak and Bax) can be activated directly following interaction with the BH3-only Bcl-2 protein Bid. Alternatively, binding of other BH3-only proteins (e.g., Noxa, Puma, Bad, and Bim) to antiapoptotic Bcl-2 proteins (e.g., Bcl-2 and Bcl-XL) results in activation of Bax and Bak. The regulated release of proapoptotic factors from the mitochondria cause induction of downstream caspases, and potential loss of mitochondrial function [28].
Fig 2.Apoptosis pathway
Cells have a discrete cell death pathway defined by a specific set of genes. These genes encode proteins that form the biochemical cascade that ultimately leads to cell death (Fig. 2). The key genes that control the cell death process are the cell death effectors of the CED-3/ICE ("caspase") family and the cell death inhibitors of the BCL2 family [29].
We probe into CED-3 and CED-6 gene expression in apoptosis pathway. In our experiment,
We suppose the magnetic field induction to apoptosis.
1.7.2 C.elegans apoptosis and cancer related gene in wormbase
Form wormbase website (Fig 3), we can find thousands of kinds of genes about the C.elegans.
Among which are about apoptosis, cancer and nerve, etc.
How could we use wormbase? Wormbase is database of an introduction C.elegans, include data of nematode genome inside, main function
(1) sequencing, genome of C.elegans has already been solved out, can find the materials of the array on this website.
(2) Mapping can find out the gene location is in C. elegans which of one chromosome at the position (3) Phenotypic information has database which lists RNAi to C. elegans
If you want to experiment as RNAi of model organism, you can consult this website.I find some genes, as apoptosis related gene, unusual function with magnetic field in C. elegans
Fig 3. Wormbase website
Chapter 2
Research experiment and design 2.1 C.elegans culture
The nematode Caenorhabditis elegans has proven to be an important model organism for.
aging-related research, providing a well characterized paradigm for NGM + 50 μM FUDR (DR-FD media) culture at 20oC
2.2 Magnetic field design
Fig 4 . Magnetic field for experiment
What we work is research that under the function of the magnetic field C.elegans biological behavior and physiological mechanism enable denying changing?
Yes .Change in our experiment.
We have designed a series of experiments and inferred for this the gene expression.
First, we buy to the strongest and steady magnet NdFeB N38 (Fig 5) on the market.
Fig 5. Manufacturer's information
Force the nematode to grow under stabilizing and covering on the function of the magnetic field with my design of the experiment. We use 6cm plate and cover and invite 3mm agar. Because the magnetic field is inversely proportional to the square of distance strongly Make use of this best magnetic field which we can obtain of method (Fig 4) .Our magnetic field that quantity examine is 0.5T on.
2.3 Video record system
We combines against Sony video cassette recorder three-dimensionally and microscope notebook computer (Fig 6). This kind of method lowers costs, and the result is very good too. In me experiment, I 8design many device that not been used before this.
Fig 6.Video record system
2.4 Experiment flow chart
Fig 7. Experiment flow chart
Magnetic design
Behavior different biochemical action
protein
2.4.1Flow chart of gene analysis
Fig 8.Flow chart of gene analysis chart
2.5 SDS-PAGE GEL
SDS-PAGE、officially sodium dodecyl sulfate polyacrylamide gel electrophoresis、is a technique used in biochemistry、genetics and molecular biology to separate proteins according to their electrophoresis mobility (a function of length of polypeptide chain or molecular weight as well as higher order protein folding、posttranslational modifications and other factors).
Fig 9.SDS-PAGE device ＆ sample
2.5.1 SDS-PAGE protocol Materials
30% acrylamide 10% SDS
10% APS (make fresh each time) TEMED
0.006% (w/v) Coomassie Blue dye 90% ddH2O
Isopropanol Fixing Solution 10% (v/v) acetic acid 25% (v/v) isopropanol 65% ddH2O
SDS sample loading buffer (40 ml) ddH2O 16 ml
0.5 M Tris, pH 6.8 5 ml 50% Glycerol 8 ml 10% SDS 8 ml
2- mercaptoethanol 2 ml (add immediately before use) bromophenol blue
10% (v/v) acetic acid Protocol
1. Prepare polyacrylamide gel according to standard protocol.
2. Load samples and run gel @ 25 mA (2 gels run @ 50 mA) in 1x SDS Running Buffer.
3. At this point, the gel can either be transferred to a membrane (see Western protocol) or stained with Coomassie (see below).
4. Place gel in a plastic container. Cover with isopropanol fixing solution and shake at room temperature. For 0.75 mm-thick gels, shake 10 to 15 min; for 1.5 mmthick gels, shake 30 to 60 min.
5. Pour off fixing solution. Cover with Coomassie blue staining solution and shake at RT for 2 hr.
6. Pour off staining solution. Wash gel with 10% acetic acid to destain, shaking at RT ON.
2.6 Primer design
The gene of interest usually has to be amplified from genomic or vector DNA by PCR (polymerase chain reaction) before it can be cloned into an expression vector. The first step is the design of the necessary primers.
Important features are:
Primer sequence. Especially the 3'-end of the primer molecule is critical for the specificity and sensitivity of PCR. It is recommemded not to have:
Primer pairs should be checked for complementarity at the 3'-end. This often leads to primer-dimer formation . Bases at the 5'-end of the primer are less critical for primer annealing. Therefore, it is possible to add sequence elements, like restriction sites, to the 5'-end of the primer molecule.
We use a primer length of 18-23 bases is optimal for our PCR applications Find out the gene that we are interested in from wormbase . Make using of knowning gene sequence to design the primers.
..
2.6.1 Gene primer designed
We look for the suitable gene correlated with our design environment as much as possible
With the behavior and observation of C.elegans, we roughly sum up into the gene that four groups are correlated with and form a cluster.Among them it is UNC family correlated with the behavior , cancer, apoptosis and anti-oxidant groups(Fig 10). Verified in the past research that there are a large amount of databases for guessing in the the genome
gpd-4
Fig 10.Gene primer classification
2.6.2 Gene primer designed list
Fwd sequence Rev sequence
unc-1 TAGAACGTGTGGAAGTGAAGG GTTGCAAATCGGAATTTGG
unc-103 CCAAGCAACTTGCACAACC GTCGTTCGAAACCTCATCG unc-112 GAAGAAGCCACACTATTTGC CACCTCCTGTTGATATGACG unc-108 GGAAAATCCTGCTTGCTCC CTCATTGTTAATGTCGAACACG unc-26 CTATTCCTCTTCGCAAGACC CCCTTCTTTCTCTGATCTTTCC unc-30 CACCATTAACCCACAATCC CAGAGTCGTCTAGCTGACTGC unc-31 AAGCGATCACTGCTCTCC CTCGAATTCTAGCCAATTCC unc-4 ACGATAACCTGTTGATGAACG CACGAATTACATTCCCACC unc-40 CACCAAGTGGAGGACAACC AAACCCAATCCGAGAAGG
unc-42 AATAGCTTCCGCCAAAGG GCAATCGAAGATCCAAATACC unc-5 CATGTACCGTTCCACCTCC CCGTTTTCCACCAGATCC
unc-51 CGAGTCAACGTTTGACCTACC TCCGTCTGCTGATACTTTGC unc-52 CCCACAAACACAGACAATCC ATTCCCCGACATTTCTATCG unc-54 CGGAAAGACTGAGAACACC ACTTCATGTTACCCATGTGC unc-58 TTATGTTTCCCCTACTCGTCC CCATCATCTGCATCGAGC
unc-59 GCAGCACAAAGAAAATCC ACACGTCCATGAAGATGC
unc-6 CCATTTCCAATCCCATCC CATCATCCGTCTCGTTTCC
unc-61 ACCGCTTTTCTCATCGTCC TCCTGTTCCCGTTTCTCC
unc-64 TCCAGTTAACGATCAGAAAACC AATTCTTTCGCGTGCTCC
unc-7 CAGACATCTCGACAATCTTCC ACTGTGCCAATACAACAATCC
unc-71 CTGCAAAAACCCAAAACC CCACACGATTTTGAATAACC unc-75 CCAAAAGACAAAATGGACG ATAAAACTGGGTTTGCTGC unc-76 CAACCTCTCGACTTTGGAACC TCATCTCGATGTGCCAGC
abl-1 CATCATCATCATCTGCACC TAGTTCCCATTCGTCTGG alx-1 TCATGCTGCTGCTAAGAAGG GCTGTTCACTGGACATTCG bath-41 ATTCACAGAGTTTCTATCCACC GCACTCATGTTTATGATTCG bath-42 CATAGGAGAATGCCCTAATCC GATATCGAGATGTTGAAGAAGC
bath-43 GACAACAAACAGTTCTCC TGCTCTAAAGACTTCTTCC
bath-44 AGAGCAGCTCAAAATACTCG CCAAACCATACATTTCAGC bir-1 GGGACCAAAAAAAAGTCG GGCTGTTAGATTTTGTTGTCG bir-2 GCTCAAAAACTTGAAAGACG GAAATCAATTTCCTGGTGG c04e12.7 GTACAAACTCACACTGTAACG AGCTTTTCCACTTCAACC
cdl-1 TCACCTTCAAAACGTCGTCC TGAGTCCACATCGAACTTTCC ced-3 TCGCTCATTCAGCAAAGC AGAATCCAAGACTGGGAATCC ced-2 GGGAACCACTGGATTAGTGC GAACAAAATGGAATGGACACG
ced-6 ACCAAACATTCCTCCATCC TCCACTTCATTGATCATTGC
ced-8 GCTCATCCAAGAAGAACTGG AATGCGACAAGTGTGATAATCC
ces-1 CTCTTCTAGTTCGTCAACGTCC ATGTGCGCTCGTAAGTTGC
che-13 TGCAAACATAATGGCTGC TATCGAGATAAGGACCAACG cps-6 CAAGCCAGATATCACTTTCC GAGCGAAAATCTCAAGACC crn-2 CGCTTCACAATTTCTCTTCG CGTTCAATGCACATGTAATCG crn-3 CTTTATCATTGATCCCTTCC AAGATGTGTAAGTGCATAGTCC csp-1 CCCACCAAACTAGATGACC CACCCATGTTCAATTTTCC
csp-2 ATATCCTGTCTGCCTTGC CTCATCCTTCTTGAATCTCG
efl-2 AAGTCAGCAATCACTTGG CATTTGTGAAGAGCAACG fem-1 ACGAAAACATTCAAGGAACACC ATTCATCGCCGACATTGC mrt-2 AGGTCTTCAAAACCGTCG ATCGAATTCAAAGTCGAGC unc-1 AAACTCTCGACGACAACG GAATATCCAGCCATAGTCTCC pik-1 CATGAAACGAGAAGTTCTCC CCAATTACATTGCTCACAGC rbx-2 CAATGCAGATTCACAAGAAGG AGAACGGGGTGAAAAATGG
t02c5.1 ATGAGTCCCCTTACGTTACC CGAAACAATTTCGCTTCC T12G3.2 CTATGATCCACCGATTCC CATCCTCTCCACTTGAACC T22H2.5 AACAAGCAATCACAACTCAGC ATCACATTTCCATCTCCATCC T27F7.2 TCATCATCTTCCACTGACACG ATTCCTCCACCACATCTTCC
tir-1 TCTCTTCTCTCCTCACTCACC TTTGCCATACATGCATCC
Y50C1A.1 AATGAAATTCGAACCCACG GGCTCCATCTTTAGGCACC
Y50E8A.9 GACACCAATCGTTCTTCC CATTTGACGAGCAAATCC
ZK1053.5 ATGTGCTCATCGTCAAACG ACAATCAAAAATGTGGCTCC ape-1 TGATCACATTTTCTCCACC TCCATTTCAAGATTCTGTCC
Fwd sequence Rev sequence
msh-2 ACTCGACCGAATCTTCTTCC CAAAATGTCCTGTGCAATAGC nft-1 CCCAACTGCTCTCATTTCC CTCCAGCATTGCTTGAACC par-4 TTTAACGAGCGTCCAACG TCTGAAGCTGAAGCAACACC
t13h5.8 TTGTTCAACCCAACCACG CAAACATGATTGGAAGGTATCC dic-1 CAACACACTTGGTTTGTGC GATCATATGCGTCGATTCC dog-1 AACGAAACACGTTCTATAGTCC AATTCCGACAGAAATCACC hoe-1 GAACATATGGACAAATGAGAGC TCAACCAACAAATCACATGG
Fwd sequence Rev sequence
maco-1 GAAATCTGAAGCTAGAGTCG GTCAGAATCATTTCCAGC cyp-44A1 CGATCTTCAGTTCAACACG GATCAAATAGCTTGCAAACC
E02H1.6 GACGACCGAATATCCTGG GTGCTCATTTTTGAACTCTCC
gcs-1 TGTTGTCTTCAAGTCACTTTCC TTTTCTCATCGTCCTGTTCG pqm-1 CAAGAAGTATATCGAACAGACG ACTGCTAAGCAGATTTTTCC
skn-1 TCCAGTTATGCCAATACTCACC TCCTTCTTCTCTTGAAACATCC
sod-2 GGAGCCTGTAATCAGTCACG TGGCGATCTTCCAAATAGC oxi-1 CCCATCACCAACATCATCC CCGAAATATTGCACATTGC E04F6.5 CGCTGAGTAAATCTGCACG AGCACCCATGACAACACC acdh-2 TGCTCAATCATCAGTCAAACC TTCCCCAAGAACGACTCC F52E1 AGCAACCAGCAAAAAGTCC CGATCCAAGAACAATGAATCC gpd-1 AGTATGACTCGACCCACG GAATGACTTTTCCCACAGC gpd-2 ACATCATCTCCAATGCTTCC TCCGAACTCGTTATCGTACC gpd-4 GATCCGTTTATCACAATCG CATCTCTCCACAGCTTTCC
Table1. Gene primer designed list
The quality and quantity of mRNA is detrimental to this experiment. Wear gloves for all procedures.
RNase is everywhere, especially on human skin. Don’t stop until reverse transcription (RT) is done.
Change gloves constantly.
2.7 RNA Concentration and Quality
The concentration and purity of total RNA are determined by measuring absorbence at 260 nm (A260) and 280nm (A280) in a spectrophotometer. The quality of total RNA is determined with agarose gel electrophoresis.
2.7.1 RNA extraction protocol
a. Homogenization, use 1 mL Tri-reagent for <0.1 gm tissue, Homogenize until the mixture turns pinky, incubate at RT 5 min, transfer 1mL mixture to 1.5mL tube
b. Phase separation, add 0.2 mL chloroform (pure), vortex, RT 2-15 min
spin, 12K rpm, 4 °C, 8 min, Immediately transfer agueous phase (~ 700 μL) to a new 2.0 mL tube
c. Column purification, Precondition
to agueous phase add equal volume (700 μL) of 70% EtOH Apply 700μL to an RNeasy mini column, spin 1 min at 12K rpm
Repeat Washing
700 μL RW1, 1 min at 12K rpm 500 μL RPE, 1 min at 12K rpm 500 μL RPE, 2 min at 12K rpm
Elution
30 μL RNAse free water, 1 min at 12K rpm Repeat, keep RNA solution on ice
Measure OD260/280 (> 2.0), in phosphate buffer (pH 7.5) (Conversion: 1 OD260 = 30 ug RNA/mL, Calculate yield)
Store unused RNA at –80 °C
Transfer 5 μL in a new tube for RT-PCR and Real-time RT-PCR.
Store the rest (45 μL) at 80 °C.
For confirming quality of RNA, we run agarose gel(Fig11) test before experiment of follow-up
Fig 11 RNA quality gel
2.8 Reverse Transcriptase from RNA to cDNA
cDNA is a more convenient way to work with the coding sequence than mRNA because RNA is very easily degraded by omnipresent RNases. The main reason cDNA is sequenced rather than mRNA. Likewise, investigators conducting DNA microarrays often convert the mRNA into cDNA
Fig 12. mRNA to cDNA
2.8.1 Reverse Transcriptase protocol
a. prepare two Annealing mixture: (2X), total RNA (10 ~30 ng)
Oligo dT(12-18) (1μg) 1μL
H2O xμL
Total 31 μL,
65 °C, 10 min, slow cool down to RT (on heat block) RT reaction,
Annealing mix 31μL 5xbuffer 10μL
0.1M DTT 5μL 2 mM dTTP 1μL Rnase inhibitor 1μL SuperScript RT II 1μL
Total 50μL
2.9 Polymerase chain reaction (PCR) principle
Polymerase chain reaction (PCR) is a simple and convenient and effective method, it can make DNA increase to above 106X in the micro- test tube. The principle of method is very simple, both ends of bit of DNA increased to want design a leading introduction respectively (forward primer) With putting the introduction instead (reverse primer) .Make it slow and cold with single whiff of goals DNA that has already changed the sex and mate (annealing) And then, utilize DNA to get ferments together (DNA polymerase) Make for the template separately by two shares of goal DNA (template) To formate new DNA share. Picture such via
(1) Denaturation, Make two shares of DNA separated. (2)It is slow and cold to mate and react (annealing), Make the introduction and goal DNA mate. (3)extension),Formate new DNA share.
Circulation operate if making by quantity of DNA each time, add every one of, if repetitive operation, many times, calculating with the mathematics formula, the quantity that DNA increases will be 2n, n is the number of times of representative's repetitive operation. In theory, one DNA is like repetitive operation PCR 20 times, then the molecule of DNA counts and increases to 220 = 106 molecule. The quantity of this DNA has been already enough to observe in agarose gel electrophoresis. [27]
Fig 13.PCR principle
2.9.1 PCR protocol
Real-time is an extension of the PCR methodology that has the potential to revolutionize the way in which PCR and quantitative PCR are conducted. In the early 1990s it was shown that the intrinsic 5＇ nuclease activity of the Taq DNA polymerase could be used to degrade specific fluorigenic probes, thereby allowing for an indirect measurement of the accumulation of the amplicon [24].
Subsequently, it was shown that using a fluorescent dye, the progress of the amplification could be monitored in a closed tube in real time using either a laser
[25] or a camera [26].
Fig 14. Real-time PCR principle
2.10.1 Relative Quantitation
One simple method for looking at relative differences in expression is the use of the relative CT approach. This approach employs the difference in the CT values obtained for two different sets of samples. Since CT values are obtained during the exponential phase of PCR, it is assumed that at each cycle the number of products are doubling. Assuming that the CT value is reflective of the initial starting copy, a difference of one.
Fluorescence systems used in Real-time-PCR. Systems employ either afluorescent dye such as SYBR Green (S) or employ a fluorescent probe that contains a reporter (R) and a quencher (Q) fluorochrome. Separation of the quencher from proximity to the reporter enables the
fluorescence of the reporter to be measured. We use SYBR Green-490 in our experiment.
Fig 15.Real-time PCR CT analyse principle
Fig 16. SYBR Green formula
2.10.2 Real-time PCR protocol
Agarose gel electrophoresis is an easy way to separate DNA fragments by their sizes and visualize them. It is a common diagnostic procedure used in molecular biological labs.
Electrophoresis:
The technique of electrophoresis is based on the fact that DNA is negatively charged at neutral pH due to its phosphate backbone. For this reason, when an electrical potential is placed on the DNA it will move toward the positive pole:
2.11.1 DNA gel protocol
1. Add the appropriate volume of 1xTAE to a sterile container 2. Add the appropriate amount of agarose
3. Heat in a microwave with the lid slightly loosened until boiling (about 2 minutes on full power) 4. Allow to cool to 'hand warm' (50-60。C) e.g. by running under a cold tap
5. Add the appropriate amount of ethidium bromide . 6. Pour the gel into the gel casting tray
7. Remove any bubbles in the gel
8. Allow the gel at least 40 minutes to set (or less if put in a fridge) ensuring that the gel casting tray is level and undisturbed
9. When the gel has set remove the combs and casting gates and transfer to the gel tank 10. Mix 5 volumes of PCR product with 1 volume of 6x gel loading buffer
11. Load samples into the wells in the gel
12. Load 1kb ladder into at least one well in each row
13. Run the gel at the appropriate voltage for about 30 minutes 14. Photograph the gel under ultraviolet light
Fig 17. electrophoresis principle
Chapr 3
Result ＆ Discussion 3.1 C.elegans behavior
Use a kind of new quantitative way in the experiment that C.elegans behavior observes Op 50 Ecoli is scribbled on agar plate, because it shed the orbits of sports when C.elegans creeps.
We use very small graph paper cushion under agar plate.We time 1 min to calculate the total length of orbit that C.elegans creep at this moment. Because we find a simple fact in the past research of our laboratory. The magnetic field could reduce the C.elegans crawling the speed.
The following charts explain observation of C.elegans' behavior
We use N2 C.elegans. We observed the C.elegansto creep and slackening. Divided into four groups as follows
C.elegans strain additive nanoparticles culture environment
N2 wild-type no 96 hours
N2+MF wild-type no 96 hours with continued magnetic field
N2+Fe3O4 Wild-type Fe3O4 6-10nm 96 hours
N2+Fe3O4+MF wild-type Fe3O4 6-10nm 96 hours with continued magnetic field
Table 2. C.elegans behavior Classification
Fig 18. C.elegans behavior with magnetic field
The above is taken a sample at random by 25, ρ＜0.05 .We find the magnetic field has lasting influence on C.elegans. The magnetic field makes the behavior slacken. Add magnetic nanoparticles increase the disparity of influencing. Assigning to four groups is for proving the prescroption and nanoparticle of the magnetic field act on together. There is an influence on influence on the magnetic field in the C.elegans for a long time
For example 2 groups 1.N2 and N2+MF
1.N2 slows down, after culturing 96 hours with continued magnetic field.
If we recorded with magnetic field, we have a fack that N2 slow down acutely . There is transient influence on influence on the magnetic field in the C.elegans during our experiment. when we take magnetic field off, the transient influence disappear. However, the long influences on slowing down still exist. The result is exciting. We add super-paramagnetic nanoparticles in our experiment. The result changes greater
3.1.1 Video recorded
We use the record system made by oneself to analyse the behavior of the C.elegans.The original text one is a video. In order to appear convenient, we use and pursue to explain in succession.
The order is 1 → 2 → 3 → 4 →5 ↓
6 → 7 → 8 →9 →10 C.elegans behavior speed
a.Control-N2＞b.N2+Fe3O4＞c.N2+magnetic field＞
d.N2+Fe3O4+magnetic field 48hr＞e.N2+Fe3O4+magnetic field 96hr a.Control-N2
Fig 19. Control-N2 mobility in succession
b.N2+Fe3O4
Fig 20. N2+Fe3O4 mobility in succession
c.N2+magnetic field
Fig 21. N2+magnetic field mobility in succession