In situ observation of the dissolution kinetics of Al_(2)O_(3) particles in CaO–Al_(2)O_(3)–SiO_(2) slags using laser confocal scanning microscopy

The dissolution kinetics of Al_(2)O_(3) in CaO-Al_(2)O_(3) SiOslags was studied using a high-temperature confocal scanning laser microscope at 1773 to 1873 K.The results show that the controlling step during the Al_(2)O_(3) dissolution was the diffusionin molten slag.It was found that the dissolution curves of Al_(2)O_(3) particles were hardly agreed with the traditional boundary layer diffusion model with the increase of the CaO/Al_(2)O_(3) ratio of slag.A modified diffusion equation considering slag viscosity was developed to study the dissolution mechanism of Al_(2)O_(3) in slag.Diffusion coefficients of Al_(2)O_(3) in slag were calculated as 2.8×10to 4.1×10m~2/s at the temperature of 1773-1873 K.The dissolution rate of Al_(2)O_(3) increased with higher temperature,CaO/Al_(2)O_(3),and particle size.A new model was shown to be v_(Al_(2)O_(3))=0.16×r_(0)^(1.58)×x^(3.52)×(T-T_(mp))^(1.11)to predict the dissolution rate and the total dissolution time of Al_(2)O_(3) inclusions with various sizes,where vAl_(2)O_(3) is the dissolution rate of Al_(2)O_(3) in volume,μm^(3)/s;x is the value of CaO/Al_(2)O_(3) mass ratio;R_(0) is the initial radius of Al_(2)O_(3),μm;T is the temperature,K;T_(mp) is the melting point of slag,K.

Real time detection of antibody-antigen interaction using a laser scanning confocal imaging-surface plasmon resonance system

A laser scanning confocal imaging-surface plasmon resonance （LSCI-SPR） instrument integrated with a wavelength-dependent surface plasmon resonance （SPR） sensor and a laser scanning confocal microscopy （LSCM） is built to detect the bonding process of human IgG and fluorescent-labeled affinity purified antibodies in real time. The shifts of resonant wavelength at different reaction time stages are obtained by SPR, corresponding well with the changes of the fluorescence intensity collected by using LSCM. The instrument shows the merits of the combination and complementation of the SPR and LSCM, with such advantages as quantificational analysis, high spatial resolution and real time monitor, which are of great importance for practical applications in biosensor and life science.

Confocal Laser Scanning Microscope Evaluation of Early Bacterial Colonization on Zirconium Oxide and Titanium Surfaces:An in vivo Study

To investigate the bacterial colonization on zirconium oxide and titanium surfaces in vivo quantitatively using a confocal laser scanning microscope （CLSM）. Ten samples of zirconium oxide ceramic and commercially pure titanium were fabricated and polished using silicon carbide abrasive paper. One sample from each group was evaluated topographic pattern under a scanning electron microscope. One sample from each group was to evaluate roughness using a profilometer. Eight volunteers were selected. The samples were cemented at the buccal surfaces of upper first molars. All samples were removed after 48 hours, immersed in SYTO-9 and propidium iodide fluorescent to stain for adherent bacteria and obseIved with CLSM. Fewer bacteria were observed in zirconia group than titanium group. However, there was no statistical difference between two groups. The experimental results demonstrate that zirconium oxide may be considered as a promising material for dental implant abutments.

EFFECT OF THE TOTAL SAPONIN OF DIPSACUS ASPER ON INTRACELLULAR FREE CALCIUM CONCENTRATION IN THE CELLULAR MODEL OF ALZHEIMER'S DISEASE-SCANNING CONFOCAL MICROSCOPY

Objective To study the effect of ginsenoside Rb1 and total saponin of dipsacus asper on intracellular free calcium concentration mediated by β amyloid protein.So as to lay a foundation for developing effective Chinese traditional medicine to treat Alzheimer’s disease.Methods The technique of laser scanning confocal microscopy combining primary cultured neurons was adopted to quantitatively analyze the change of [Ca 2+ ] i.Results The [Ca 2+ ] i of primary cultured hippocampal neurons was nmol·L -1 on basal levels.Control group showed obvious change of calcium vibration,[Ca 2+ ] i was elevated to nmol·L -1 .The peak of [Ca 2+ ] i of Rb1 group reached nmol·L -1 and was lower than that of control group .The tSDA group displayed distinct change of calcium vibration too,and [Ca 2+ ] i reached nmol·L -1 .There was a significant difference in [Ca 2+ ] i between control and tSDA group .Conclusion The research indicated that one of mechanisms by which Rb1 and tSDA protected the neurons was to maintain the balance of [Ca 2+ ] i.

Quantum Dots as Fluorescent Labels for Detection of Heat Shock Protein in Tumor Tissue Using Laser Scanning Confocal Microscope

A new Quantum Dots（Qdots） nanocrystal composed of semiconductor core and zinc sulfide shell, and its feasibility as labels in immunofluorescence analysis for the imaging of tumor biomarkers by laser scanning confocal microscope（LSCM） was investigated. Qdots taged by mercaptoacetic acid were conjugated with second antibody, then imaging differences of Heat Shock Proteins 70（HSP70） in renal carcinoma tissure sections with immunofluorescence analysis method using Qdots bioconjugates and conventional organic dye FITC were observed by LSCM to assess the brightness and opticalstability of Qdots. The experimental results showed Qdots bioconjugates achieved the better results in demonstrating HSP70 with more brighter color and more clear picture than FITC labels. Moreover, the label signals of Qdots did not fade clearly after continued exposure to a 488 nm laser for 1 h. The Qdots bioconjugates have good feasibility in immunofluorescence analysis for the bioimaging by LSCM.

Dissolution behavior of Al_(2)O_(3)inclusions into CaO-MgO-SiO_(2)-Al_(2)O_(3)-TiO_(2)system ladle slags

To investigate the dissolution behaviors of Al_(2)O_(3)inclusions in CaO-5wt%MgO-SiO_(2)-30wt%Al_(2)O_(3)-TiO_(2)system ladle slags,confocal scanning laser microscopy was conducted on the slags with different TiO_(2)contents(0-10wt%),and scanning electron microscopy was performed to study the interfacial reaction between Al_(2)O_(3)and this slag system.The results disclose that the dissolution of Al_(2)O_(3)inclusions does not result in the formation of new phases at the boundary between the slag and the inclusions.In TiO_(2)-bearing and TiO_(2)-free ladle slags,there is no difference in the dissolution mechanism of Al_(2)O_(3)inclusions at steelmaking temperatures.Boundary layer diffusion is found as the controlling step of the dissolution of Al_(2)O_(3),and the diffusion coefficient is in the range of 4.18×10^(-10)to 2.18×10^(-9)m^(2)/s at 1450-1500℃.Compared with the solubility of Al_(2)O_(3)in the slags,slag viscosity and temperature play a more profound role in the dissolution of Al_(2)O_(3)inclusions.A lower viscosity and a lower melting point of the slags are beneficial for the dissolution.Suitable addition of TiO_(2)(e.g.,5wt%)in ladle slags can enhance the dissolution of Al_(2)O_(3)inclusions because of the low viscosity and melting point of the slags,while excessive addition of TiO_(2)(e.g.,10wt%)shows the opposite trend.

Highly specific characterization and discrimination of monosodium urate crystals in gouty arthritis based on aggregation-induced emission luminogens

Existing technologies used to detect monosodium urate(MSU)crystals for gout diagnosis are not ideal due to their low sensitivity and complexity of operation.The purpose of this study was to explore whether aggregation-induced emission luminogens(AIEgens)can be used for highly specific imaging of MSU crystals to assist in the diagnosis of gout.First,we developed a series of luminogens(i.e.,tetraphenyl ethylene(TPE)-NH_(2),TPE-2NH_(2),TPE-4NH_(2),TPE-COOH,TPE-2COOH,TPE-4COOH,and TPE-Ketoalkyne),each of which was then evenly mixed with MSU crystals.Next,optimal fluorescence imaging of each of the luminogens was characterized by a confocal laser scanning microscope(CLSM).This approach was used for imaging standard samples of MSU,hydroxyapatite(HAP)crystals,and mixed samples with 1:1 mass ratio of MSU/HAP.We also imaged samples from mouse models of acute gouty arthritis,HAP deposition disease,and comorbidities of interest.Subsequently,CLSM imaging results were compared with those of compensated polarized light microscopy,and we assessed the biosafety of TPE-Ketoalkyne in the RAW264.7 cell line.Finally,CLSM time series and three-dimensional imaging were performed on MSU crystal samples from human gouty synovial fluid and tophi.As a promising candidate for MSU crystal labeling,TPE-Ketoalkyne was found to detect MSU crystals accurately and rapidly in standard samples,animal samples,and human samples,and could precisely distinguish gout from HAP deposition disease.This work demonstrates that TPE-Ketoalkyne is suitable for highly specific and timely imaging of MSU crystals in gouty arthritis and may facilitate future research on MSU crystal-related diseases.

IN SITU OBSERVATION OF GROWTH BEHAVIOR AND MORPHOLOGY OF DELTA-FERRITE AS FUNCTION OF SOLIDIFICATION RATE IN AN AISI304 STAINLESS STEEL

It was presented the in situ observation of growth behavior and morphology of delta-ferrite as a function of solidification rate in an AISI304 stainless steel. The specimens have been solidified and observed using confocal scanning laser microscopy (CSLM). The δ-phase always appears like cells on the sample surface when critical supercooling occurs, during which the L→δ transformation starts. The solid-liquid (S-L) interface is found to be finger shaped and has no faceted shape. γ phase appears among δ grains due to partitioning of Ni into the melt during solidification, when solidification rate is higher. The mergence of observed δ cells is possible for the steel sample cooled at 7.5℃/min. The formation of dendrites can be observed on the free surface of the steel sample cooled at 150℃/min. The size of solidified delta grains decreases from 120 to 20-80μm, and the volume fraction of solidified austenite increases with increase in solidification rate from 7.5 to 150℃/min. The relation between the tip radius of δ cell and its growth rate is deduced, and the results agree with the experimental values.

Retinal displacement after closure of idiopathic macular hole

Objective: To study the foveal displacement during the closure of idiopathic macular holes(MHs).Methods: Thirty-seven idiopathic MH patients treated by pars plana vitrectomy and internal limiting membrane peeling were studied prospectively.Locations of MH center and foveal pit were measured by optic coherence tomography.Retinal displacement was observed using confocal scanning laser ophthalmoscopy.Results: A total of 40 eyes were included in this study and MHs were closed in 37 eyes(92.5%).The confocal scanning laser ophthalmoscopy showed that all of the retinal capillaries in the superior, inferior, nasal and temporal sides of the MHs moved toward the optic nerve head(ONH).The optic coherence tomography results showed that the mean nasal displacements of foveal pits were(102.9±61.2),(109.6±53.1), and(137.0±52.0) μm at 3, 6 and 12 months, respectively.And the mean vertical displacements were(55.9±49.4),(61.4±57.8) and(67.8±54.3) μm, respectively.Post-operative foveal pits were located in the nasal side of the MH centers.The extension of retina and nasal to the MH were in opposite directions: the nasal hole margin moved toward the MH, but the retina located closer to the ONH moved toward the ONH.The fellow eyes of three patients developed into idiopathic MH during the follow-up period and operations were performed for all of the three patients.Conclusion: Our results showed that center of macula does not move when an idiopathic MH develops, but it moves toward ONH during closure of hole; thus, new fovea is in nasal side of original fovea.

Changes of meibomian glands in patients with type 2diabetes mellitus

AIM： To investigate the morphological changes of meibomian glands in patients with type 2 diabetes mellitus （DM）.METHODS： Of 118 eyes （118 patients） with type 2 DM （DM group） and 100 eyes of 100 control subjects （control group） were enrolled. After completing an ocular surface disease index （OSDI） questionnaire, the non-invasive tear film break-up time （NI-BUT） and the structure of the meibomian glands （MGs, meibography） were assessed by the Keratograph 5M system. Partial or complete loss of MG was scored for each eyelid from grade 0 （no loss） to grade 3 （lost area was 〉2/3 of the total MG area）, which were also examined by laser scanning confocal microscopy （LSCM）. The primary outcomes were meibomian gland acinar unit density （MGAUD）, meibomian gland acinar longest diameter （MGALD） and meibomian gland acinar shortest diameter （MGASD）.RESULTS： Compared with control group, the OSDI was significantly higher in DM group （Z=-5.916; P〈0.001）, while the NI-BUT was significantly lower （Z=-7.765; P〈0.001）. Keratograph showed that there were more MGs dropout in DM group than that in control group. The meiboscore was significantly higher in DM group compared with control group （Z=-3.937; P〈0.001）. LSCM revealed that there were cytological alterations of MGs in DM group compared with control group, which included enlargement of MG acinar units and decreased in density of MG acinar units. Specifically, there were lower MGAUD, larger MGALD and MGASD in DM group than control group （Z=-10.120, -9.4442, -7.771; P〈0.001）.CONCLUSION： Compared with the normal control participants, the patients with type 2 DM had more unstable tear films and severe symptoms of dry eye. Using Keratograph 5M system and LSCM, we found that the patients with type 2 DM had more significant morphological and cytological changes and dysfunction in MGs.

Effect of melatonin on the spatial and temporal changes of [Ca^(2+) ]i in single living cells of cortical neurons by laser scanning confocal microscopy

Objective To examine the effects of melatonin on the dynamic changes in the concentration of intracellular free Ca 2+ ([Ca 2+ ]i) in single intact cultured cortical neurons isolated from fetal rats, in order to explore the possible antiaging mechanisms of melatonin (MT) Methods Using the highly fluorescent Ca 2+ sensitive indicator Fluo 3/AM, cortical neurons cultured in a 35?mm Tissue Culture Dish were in incubated for 45?min at room temperature with 5?μmol/L Fluo 3/AM, resulting in proper intracellular dye concentration to provide adequate signal strength for detection and excellent Laser Scanning Confocal Microscopy (LSCM) imaging of [Ca 2+ ]i while not disturbing normal intracellular physiology The changes in fluorescent intensity were monitored by LSCM Results Bay K8644 (10 6 ?mol/L), KCl (20 ?mmol/L), sodium L glutamate (Glu, 50?μmol/L) caused a rapid increase of [Ca 2+ ]i in cortical neurons, and this increase could be significantly attenuated by 10 6 and 10 7 mol/L MT Conclusions MT could antagonize the extracellular Ca 2+ influx, reduce Ca 2+ overload, and have a protective effect on neurons This may be one of the important antiaging mechanisms of MT

Endosperm Development in Autotetraploid Rice

By using the laser scanning confocal microscope and plastic （Leica 7022 historesin embedding kit） semi-thin sectioning technique, comparative studies on the development of endosperm were carried out between autotetraploid and diploid rices. About one third of the ovaries in the autotetraploid showed normal endosperm development as those in the diploid. In these ovaries, one of the polar nuclei would fuse with the sperm nucleus, and the primary endosperm nucleus formed and underwent the first division in 4 hours after pollination; the anticlinal wall began to grow centripetally between the free nuclei starting from the wall ingrowths of the embryo sac near the micropylar end, and some of the phragmoplasts formed transformed into periclinal walls. In addition, some of the cell wall situated in the middle of the endosperm appeared to originate from phragmoplasts, whereas others seemed to develop randomly without the obvious formation of phragmoplasts. Cellulose began to accumulate in the wall of aleurone cell layer at 6 days after pollination. The cellulose wall of the cells of the aleurone cell layer appeared to have completely formed within 7 to 8 days after pollination. On the other hand, about two thirds of the ovaries in the autotetraploid showed abnormality in endosperm development with various types, such as non-fertilization, abnormal fertilization, endosperm development-delay and non-synchronization in the development of cellulose wall of cells of the aleurone layer. These abnormalities usually resulted in decreased seed setting in autotetraploid rice.

Effect of Cholic Acid on Fetal Cardiac Myocytes in Intrahepatic Choliestasis of Pregnancy

This study examined the effect of cholic acid （CA） on cultured cardiac myoeytes （CMs） from neonatal rats with an attempt to explore the possible mechanism of sudden fetal death in intra- hepatic cholestasis of pregnancy （ICP）. Inverted microscopy was performed to detect the impact of CA on the beating rates of rat CMs. MTT method was used to study the effect of CA on the viability of CMs. CMs cultured in vitro were incubated with 10 ~maol/L Ca2＋-sensitive fluorescence indicator fluo-3/AM. The fluorescence signals of free calcium induced by CA were measured under a laser scanning confocal microscope. The results showed that CA decreased the beating rates of the CMs in a dose-dependent manner. CA could suppress the activities of CMs in a time- and dose-dependent manner. CA increased the concentration of intracellular free calcium in a dose-dependent manner. Our study suggested that CA could inhibit the activity of CMs by causing calcium overload, thereby leading to the sudden fetal death in ICP.

Lead Can Inhibit NMDA-,K^+-,QA/KA-Induced Increases in Intracellular Free Ca^(2+) in Cultured Rat Hippocampal Neurons

Objective To examine the effects of Pb2+ on N-methyl-D-aspartate (NMDA)-, K+- and quisqualate(QA)/kainite(KA)-induced increases in intracellular free calcium concentration ([Ca2+],) in cultured fetal rat hippocampal neurons in order to explain the cognitive and learning deficits produced by this heavy metal. Methods Laser scanning confocal microscopy was used. Results The results clearly demonstrated that adding Pb2+ before or after NMDA/glycine stimulation selectively inhibited the stimulated increases in [Ca2+], in a concentration-dependent manner. In contrast, Pb2+ treatment did not markedly affect increases in [Ca2+], induced by an admixture of QA and KA. The minimal inhibitory effect of Pb2+ occurred at 1 μmol/L, and more than seventy percent abolition of the NMDA-stimulated increase in [Ca2+]; was observed at 100 Jμmoll/L Pb2+. Evaluation of Pb2+-induced increase in [Ca2+], response to elevating extracellular concentrations of NMDA, glycine or calcium revealed that Pb2+ was a noncompetitive antagonist of both NMDA and glycine, and a competitive antagonist of Ca2+ at NMDA receptor channels. In addition. Pb2+ inhibited depolarization-evoked increases in [Ca2+], mediated by K+ stimulation(30μmol/L). indicating that Pb2+ also depressed the voltage-dependent calcium channels. Also, the results showed that Pb2+ appeared to be able to elevate the resting levels of [Ca2+|, in cultured neurons, implying a reason for Pb2+-enhanced spontaneous release of several neurotransmitters reported in several previous studies. Conclusion Lead can inhibit NMDA-. K+-, QA/KA-jnduced increases in intracellular [Ca2+], in cultured hippocampal neurons.

Single Particle-Based Confocal Laser Scanning Microscopy for Visual Detection of Copper Ions in Confined Space

Main observation and conclusion A single particle-based confocal laser scanning microscopy was developed for the visual detection of copper ions in confined space.A fluorescence microparticle,named AuNCs/ZIF-8,was synthesized by coating gold nanoclusters(AuNCs)onto the outer surface of zeolitic imidazolate framework-8(ZIF-8).

Overexpression of Myocardin-related transcription factor-A attenuated middle cerebral artery occlusion/reperfusion-induced apoptosis via the Mcl-1/Cyt C/cleaved caspase 3 pathway

To investigate the effect of Myocardin related transcription factor A(MRTF-A)on apoptosis induced by ischemic/reperfusion(I/R),middle cerebral artery occlusion/reperfusion(MCAO/R)in rats were applied to mimic I/R.The neurological deficit score,cerebral infarct size,cortical neuron apoptosis and cleaved caspase 3 level were evaluated to determine the effect and the level of apoptosis by TTC straining,terminal deoxynucleotidyl transferase dUTP nick end labeling(TUNEL)straining,Western blot and immunofuorescence staining.The myeloid cell leukemia-1(Mcl-1)expression,release of cytochrome C(Cyt C)and its colocalization with apoptotic pro-tease activating factor-1(Apaf-1)were analyzed by quantitative real-time PCR(qRT-PCR),Western blot,and immunofuorescence staining.The results showed that MRTF-A over-expression could decrease the neurological deficit score and reduce cerebral infarct size(P<0.01 versus Sham).In the MRTF-A-I/R group,TUNEL-positive cells and apoptosis ratio(%)(51.61±6.17%)were significantly decreased compared to the Neg-I/R group(76.45±8.77%)at 24 h reperfusion.Meanwhile,the cleaved caspase 3 expression revealed a similar trend while the expression of Mcl-1 was the opposite.Moreover,MRTF-A overexpression significantly enhanced Mcl-1 fAuorescence intensity,which up-regulated the mRNA and protein level(P<0.05or P<0.01 versus Neg-I/R).Furthermore,MRTF-A overexpression markedly inhibited the release of Cyt C,and decreased the colocalization with Apaf-1 in the cytoplasm(P<0.05 or P<0.01.versus Neg-I/R).All the data indicated that MRTF-A overexpression could improve the neu-rological function against cerebral I/R-induced apoptosis since underlying mechanism might be involved in the Mc-1/Cyt C/cleaved caspase 3 signaling pathway.

The Expression of the IGF Family During Mouse Mammary Gland Development

This study was to determine the patterns and levels of IGF family members＇ expression during postnatal mammary gland development. The authors investigated the protein expression profile of the major components of the IGF axis in murine mammary glands. All the proteins examined, IGF- Ⅰ, IGF- Ⅱ, and IGF- Ⅰ receptor （IGF-Ⅰ R） were expressed at greatly different levels and displayed unique expression profiles. IGF- Ⅱ and IGF- ⅠR were always expressed at significantly higher levels than IGF- Ⅰ. IGF- Ⅰwas localized in adipocytes as well as the epithelial and stromal compartments, but just distinctly expressed where mammary cells aggregated to form ducts, in virgins. The IGF- Ⅱ was localized only on the basal layer epithelial cell membranes of ducts and alveoli, with a peak level on the initiation of lactation. The higher level of IGF- ⅠR compared with IGF- Ⅰ was also found in adipocytes as well as in the epithelial and stromal compartments, especially during pregnancy and late lactation. The IGF- Ⅰ R pathway was obviously significant for the development of the mammary parenchyma and stroma. Overall, the comparison of the expression profiles of these different proteins would strongly suggest that they were likely to have different functions throughout the mammary gland development, and it also highlighted the potential interactions and coregulation of the members of this axis. It seems that IGF- Ⅱ was the major local modulator rather than IGF- Ⅰ by an IGF- Ⅰ R-independent pathway, especially for initiation of lactation. This study has demonstrated the importance and complexity of the IGF axis during mammary gland development and provides a valuable resource for future research in this area.

Thermo-Calc calculations and studies of the heating process of Fe_(83)Si_4B_(13) alloy

The microstructural changes of Fe83Si4B13 amorphous mother alloy during the heating process were investigated by Laser Scanning Confocal Microscopy （LSCM） ,and the phase transformation was determined by the Thermo-Calc calculations. The differences in the melting points measured by Differential Scanning Calorimetry （DSC） and LSCM, and those obtained by Thermo-Calc calculations were also discussed. It is found that the melting points measured by DSC and LSCM are relatively similar, whereas the onset and end of the melting temperatures calculated by Thermo-Calc software are higher than those measured by DSC and observed by LSCM.

Megasporogenesis and Megagametogenesis in Autotetraploid Indica/Japonica Rice Hybrid

Autotetraploid indic, a/japonica rice hybrid combines both the advantages of polyploidy and the heterosis between indica and japonica rice. Embryo sac abortion is an important factor influencing spikelet fertility in autotetraploid dce hybrid. To clarify the cytological mechanism of embryo sac abortion, the megasporogenesis and megagametogenesis in an autotetraploid japonicaAndica hybrid were examined by the whole-mount eosin B-staining confocal laser scanning microscopy （WE-CLSM） technique. Abnormalities were observed from the megasporocyte stage to the mature embryo sac stage. The degeneration of the tetrad cells and the functional megaspore was the characteristic of abnormalities during megasporogenesis. Abnormal small embryo sacs and disordered number of nuclei were frequently observed during embryo sac development. Some interesting phenomena, such as two functional megaspores, the diplospory-like megasporocyte, and five-nucleate embryo sac were found. The abnormalities that occurred during female gametophyte development resulted in more than five types of abnormal embryo sacs （i.e. embryo sac degeneration, embryo sac without female germ unit, embryo sac without egg apparatus, embryo sac with abnormal polar nuclei and abnormal small embryo sac） in autotetraploid japonica/ndica hybdd. Embryo sac fertility was lower in diploid japonica/ndica hybdd than in autotetraploid japonicaAndica hybrid although many abnormal phenomena were observed in autotetraploid hybrid.

APOPTOSIS OF HYPERPLASIA AND CANCER OF THE GALLBLADDER WITH CALCULAS

Objective To investigate the relation between different extent of proliferation caused by gallstone and gallbladder cancer by counting the proliferation and the apoptosis of the gallbladder cancer for the clinically prevention of the gallbladder carcinoma.Methods The TUNEL method was used to detect the apoptosis of the specimens and the mean apoptosis indices obtained by quantification of apoptosis cells fluorescence by laser scanning confocal microscope were compared among the variable pathological patterns.Results The mean apoptosis indexes in the mormal and abnormal specimens with cholecystitis,simple hyperplasia, low-grade dysplasia, mid-grade dysplasia, high-grade dysplasia and carcinoma were 5.11, 5.49, 6.32, 8.65, 12.27, 25.24, 39.62, 119.8, respectively. There was significant difference among the variable pathological patterns and as the lesion progressing, the index went up gradually with the carcinoma had the highest index. Conclusion The apoptosis indexes increase with the pathological progress during the carcinogenesis of gallbladder cancer caused by lithiasis, which stimulate the epithelium for long time and result in an increasing of the apoptosis; and it may play an important role in the carcinogenesis of gallbladder cancer.