CN102589947B - Preparation method of madrepore skeleton sample for being suitable for observing skeleton minute structure and researching environment record of madrepore - Google Patents
Preparation method of madrepore skeleton sample for being suitable for observing skeleton minute structure and researching environment record of madrepore Download PDFInfo
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- CN102589947B CN102589947B CN201210039382.XA CN201210039382A CN102589947B CN 102589947 B CN102589947 B CN 102589947B CN 201210039382 A CN201210039382 A CN 201210039382A CN 102589947 B CN102589947 B CN 102589947B
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Abstract
The invention relates to a preparation method of a madrepore skeleton sample for being suitable for observing a skeleton minute structure and researching an environment record of the madrepore. The preparation method comprises the steps of: (1), soaking the madrepore skeleton sample in bleaching solution for the first time, cleaning the madrepore skeleton sample by clear water, and soaking the madrepore skeleton sample in another bleaching solution for the second time; wherein the bleaching solution is 10% hydrogen peroxide with the pH value of 8-9 or NaClO (sodium hypochlorite) solution; (2), further cleaning the madrepore skeleton sample after the soaking is finished, and drying the madrepore skeleton sample at the temperature of 45-60 DEG C for using. According to the preparation method, coral polyp tissue and other organic impurities are removed by using common reagent such as the hydrogen peroxide and the like and an ultrasonic cleaning instrument, thereby obtaining clean madrepore skeleton sample, so that the damage to the skeleton minute structure and the skeleton minute structure in the preparation process is reduced to a maximum limit, meanwhile, the changes of the trace element content and component ratio of isotopes in the skeleton are avoided.
Description
Technical field
The invention belongs to biomass geochemistry field and environmental ecology field, relate to a kind of experiment by the preparation method who makes reef coral bone sample.
Background technology
Make reef coral and in vital movement process, secrete and deposit calcium carbonate, form continuous bone growth layer.Environmental change reaction is responsive to external world to make reef coral, change the diurnal periodicity of the envirment factors such as temperature, salinity, pH and all can cause the even change of health status of coral polyp physiological situation, the aragonite that its same period, calcification formed also can be affected in biological chemistry proterties, such as its contained isotope δ
18o, δ
13c, δ
11the change of B composition ratio, the content of geochemical elements (comprising heavy metal element) changes, and skeleton density band changes etc.So, make reef coral bone growth synchronous recording coral growth history and environmental change information, be widely used in biomass geochemistry, the fields such as ecologic environment, such as the even research of yield-power of palaeoclimatic reconstruction, sea level fluctuation, volcanic eruption, continent runoff, upward flow, marine pollution.Also be widely used classification and evolution, modern times of tetracoral in time immemorial of the simultaneously observation based on making reef coral skeleton and microtexture feature are made the research field such as the classification of reef coral (zoantharian) and the studies and clinical application of medical domain coral hydroxyapatite artificial bone.Before carrying out the experiment such as the isotope analysis of cave coral bone sample, Analysis of Heavy Metal, microtexture observation, all must process by early stage removal anthozen polyp soma, and contained a small amount of organic matter and inorganic carbonate cementing matter in bone.These impurity are all enough to affect the result of coral bone geochemical survey and the observation of microtexture.Therefore, it is the prerequisite of carrying out the researchs such as cave coral environment record that preparation is applicable to observe the cave coral bone sample that bone microtexture and geochemistry measures, and has great importance.
In many documents that relates to the pre-treatment of cave coral bone, have both at home and abroad quite a few do not mentioned concrete disposal route or just simply rinse, boil with fresh water or ultrasonic oscillation.These methods often cannot be removed the organic matters such as coral tissue effectively, and often can cause that coral skeletal injury even makes a variation.Widely used is corrosion and oxidizing process, can be divided into NaOH, H according to its chemical substance
2o
2, tri-kinds of different immersion process of NaClO.But not very careful about the narration of these three kinds of methods, not affecting bone microtexture for how effectively removing coral tissue and bones impurity and constituent does not have systematic discussion and introduction.In fact, the oxidisability of NaClO is the strongest, but the secondary pollution causing after its toxicity and oxidation is inevitable; Though and the corrosivity of NaOH tool excellence, but also can cause secondary pollution and disturb the observation of microtexture to only have and clean the above-mentioned impact of just preventing by enough later stages.Hydrogen peroxide, due to the singularity of its chemical composition, can substantially not have secondary pollution in being oxidized decontamination, but is faintly acid because it is water-soluble, can cause the dissolving of the micromechanisms such as aragonite needle.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, provide a kind of applicable to observing bone microtexture and studying the cave coral bone sample preparation methods of its environment record, meet different experiments and require the method for cave coral bone by the preparation of simple laboratory equipment.
Invention is achieved through the following technical solutions above-mentioned purpose:
Cave coral bone sample preparation methods of the present invention comprises the following steps: soak cave coral bone sample for the first time in bleaching liquid (1), with clear water flushing, then with soaking for the second time in new bleaching liquid; Described bleaching liquid is that pH is 10% hydrogen peroxide or the NaClO solution of 8-9; (2) after immersion finishes, cave coral bone sample is further cleaned, in 45 ℃ ~ 60 ℃ dry for standby.
In step (1), described soak time is for the first time about 1-2 hour, makes most of biological tissue and organic oxidized and depart from coral bone.Rinse 1 minute with clean fresh water subsequently, to remove the dirt of organizing after oxidized.After soaking for the first time, can produce a large amount of bubbles and historrhexis's thing, soak for the second time therefore need to change bleaching liquid.Soak time is about 24 hours for the second time,, the impurity such as the organism of further fully oxidation bones remnants.
The further cleaning that step (2) is described, is first to rinse and wash with clear water, rinses to wash step and need soft minimizing to collide with, then is placed in the low and middle-grade cleaning of ultrasonic oscillation instrument 3-5 minute.Sonic oscillation scavenging period and number of times can be taken the circumstances into consideration increase and decrease according to actual conditions, and the single concussion time can not be excessively of a specified duration, prevents water temperature over-high.After sonic oscillation, clean with deionized water again, then dry.
The bleaching liquid that the present invention adopts can adopt hydrogen peroxide solution or sodium hypochlorite (NaClO) solution, but in view of sodium hypochlorite toxicity and may produce secondary pollution, if the follow-up mensuration for trace element (comprising metallic element) of sample does not advise using; Can use liquor natrii hypochloritis if follow-up for isotopic mensuration, but must clean up.Preferably hydrogen peroxide solution, is more preferably pH and is 8.5 10% hydrogen peroxide solution.When the pH value of liquid is drifted in adjusting, can adopt NaOH to regulate.
If be subsequently applied to the mensuration of heavy metal, after sonic oscillation, cave coral bone sample further should be soaked to 10-20 minute with metal-chelating agent solution, clean once thoroughly to remove remained on surface metal and to use deionized water to repeat concussion.Metal-chelating agent solution used contains 17 mM sodium diethyldithiocarbamates, 0.4 M vitamin C and 11.7 mM lime chloride (calcium chloride); PH value is 5.Rinse after washing immersion and can take different disposal method by requirement of experiment through deionized water.If bone sample is follow-up for isotopic mensuration, bake out temperature can not exceed 45 ℃; Other purposes can be selected 60 ℃ of oven dry as one sees fit.
Compared with prior art, the present invention has following beneficial effect:
1. the present invention removes anthozen polyp soma and other organic impurities by the common reagent such as application hydrogen peroxide and ultrasonic washing instrument, thereby obtain clean cave coral bone, reduce to greatest extent the destruction of coral skeleton and microtexture in preparation process, avoided the change of micronutrient levels and isotopics ratio in bone simultaneously.
Embodiment
Further explain the present invention below in conjunction with embodiment, but embodiment does not limit in any form to the present invention.
embodiment 1
Gather nose shape deer horn from Coral Reef Region, surrounding waters, Tropical Ocean Bioexperiment station, South Sea institute of oceanography of the Chinese Academy of Sciences 3 m depth of waters
pocillopora damicornis.Intercepted length is that the coral sample of 5cm is placed in 10% hydrogen peroxide solution that adjusted pH value is 8.5 and soaks 2 hours.Take out coral bone tap water and rinse gently the hydrogen peroxide solution immersion 24 hours that reuses same concentrations and pH value after washing.Rinse wash clean with distilled water, then use ultrasonic oscillation 5 minutes, finally with deionized water soak rinse wash after in 60 ℃ of oven dry of baking oven.For the observation of bones structure and microtexture.
Complete through above-mentioned steps coral bone surface after treatment and inner structure preservation, after 4 years, the secondary aragonite structure in interseptal space still retains intact.
embodiment 2
Gather the cup-shaped coral of deer horn from Coral Reef Region, marine site, Xi Mao island, the Sanya 3m depth of water
pocillopora damicornis, the coral of 10 3cm of intercepting, is placed in 10% hydrogen peroxide solution that adjusted pH value is 8.5 and soaks 2 hours.Take out coral bone tap water and rinse gently the hydrogen peroxide solution immersion 24 hours that reuses same concentrations and pH value after washing.Rinse wash clean with distilled water, use Ultrasonic Cleaning 10 minutes, be then soaked in chelate remover 10 minutes, rinse after washing and reuse Ultrasonic Cleaning 10 minutes with deionized water.Finally in 60 ℃ of oven dry of baking oven.For the mensuration of bone heavy metal (Cu, Pb, Cd, Zn, Ni).
Complete through above-mentioned steps coral bone surface after treatment and inner structure preservation, the secondary aragonite structure after 3 years in interseptal space retains intact.
Claims (7)
1. one kind applicable to observing bone microtexture and studying the cave coral bone sample preparation methods of its environment record, it is characterized in that comprising the following steps: soak cave coral bone sample for the first time in bleaching liquid (1), rinse with clear water, then soak for the second time with new bleaching liquid; Described bleaching liquid is that pH is 10% the hydrogen peroxide of 8-9 or pH be 8-9 10% NaClO solution; (2) after immersion finishes, after cave coral bone sample is first rinsed and washed with clear water, then be placed in ultrasonic oscillation instrument, the low-grade 3-5 minute that cleans, again cave coral bone sample is further soaked to 10-20 minute with metal-chelating agent solution, in 45 ℃～60 ℃ dry for standby.
2. the method for claim 1, is characterized in that the described soak time for the first time of step (1) is 1-2 hour.
3. the method for claim 1, is characterized in that the described soak time for the second time of step (1) is 24 hours.
4. the method for claim 1, is characterized in that described the flushing with clear water of step (1) is to rinse 1 minute with clean fresh water.
5. the method for claim 1, is characterized in that, before the described oven dry of step (2), also bone sample being cleaned with deionized water.
6. the method for claim 1, is characterized in that hydrogen peroxide or NaClO solution that step (1) is described are to regulate pH value to 8.5 with NaOH.
7. the method for claim 1, is characterized in that the described metal-chelating agent solution of step (2) contains 17mM sodium diethyldithiocarbamate, 0.4M vitamin C and 11.7mM lime chloride (calcium chloride); PH is 5.
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| CN108548826A (en) * | 2018-04-08 | 2018-09-18 | 广西大学 | A method of differentiating that artificial seawater cultivation coral and natural waters grow coral |
| CN109738248A (en) * | 2018-12-25 | 2019-05-10 | 海南热带海洋学院 | A kind of spicule extracting method of meat sesame soft coral |
| CN110006728A (en) * | 2019-03-20 | 2019-07-12 | 中国地质大学（武汉） | A method of efficiently separating clast thorium in offshore coral bone |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110185946A1 (en) * | 2008-10-09 | 2011-08-04 | Metabiomed. Co. Ltd | Porous composite comprising silicon-substituted hydroxyapatite and ß- tricalcium phosphate, and process for preparing the same |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| US20110185946A1 (en) * | 2008-10-09 | 2011-08-04 | Metabiomed. Co. Ltd | Porous composite comprising silicon-substituted hydroxyapatite and ß- tricalcium phosphate, and process for preparing the same |
Non-Patent Citations (7)
| Title |
|---|
| Pre-treatment effects on coral skeletal δ13C and δ18O;A. G. Grottoli et. al.;《Chemical Geology》;20051231;第221卷;第225-242页 * |
| Pretreatment of coral aragonite for Mg and Sr analysis: Implications for coral thermometers;T. Watanabe et. al.;《Geochemical Journal》;20011231;第35卷;第265-269页，尤其是第266页左栏第2段8-10、14-16、19行 * |
| 三亚活体珊瑚的微量元素与硼同位素组成的初步研究;李华玲等;《盐湖研究》;20060630;第14卷(第02期);第35-41页 * |
| 介形虫碳氧同位素测定样品处理方法对比研究;李祥忠等;《盐湖研究》;20070331;第15卷(第01期);第5-11页，尤其是第7页左栏第1段 * |
| 天然滨珊瑚无害化处理后形态学和抗压强度的变化;董青山等;《临床口腔医学杂志》;20090630;第25卷(第06期);第338-340页 * |
| 正热电离静态多接收偏硼酸铯离子测定硼同位素;贺茂勇等;《分析化学》;20090531;第37卷(第05期);第703-706页，尤其是第704页2.2 * |
| 珊瑚中硼的分离及其同位素组成的测定;张崇耿等;《理化检验.化学分册》;20031130;第39卷(第11期);第652-654页 * |
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