Further, since MPL is a potent inducer of

Further, since MPL is a potent inducer of Seliciclib in vivo Th1 response and can function through subcutaneous route also, we speculate that MPL can be combined with liposomes and can be administered through subcutaneous route to overcome the failure of liposomal vaccine through this route. Indeed we have preliminary evidence showing

that immunization with liposomal antigens in association with MPL-TDM can induce protection against L. donovani infection in BALB/c mice through subcutaneous route (unpublished observation). AS01, a liposomal formulation containing MPL as a potent inducer of humoral and cell-mediated response is already in clinical trials for malaria [10]. Thus liposomal formulated MPL-TDM+LAg may be the choice of adjuvant for vaccine development against Leishmania and other intracellular pathogens. Conclusions This

comparative study of BCG+LAg and MPL-TDM + LAg vaccines with cationic liposomal formulation of LAg interestingly reveals a significantly greater effectiveness of the liposomal vaccine for protection against progressive VL in BALB/c. Evaluation of the immune responses emphasize the need for an immunogenic vaccine for elicitation of potent vaccine-induced cellular immunity based on both Th1 and Th2 cell responses to confer protection against the visceral disease. Thus, the cationic liposomes offer a rational choice of adjuvant for the development of vaccines against a range of infectious diseases such as LBH589 supplier leishmaniasis, malaria and tuberculosis. Methods Animals Female BALB/c mice (4-6 weeks old),

bred in the animal facility of Indian Institute of Chemical Biology (Kolkata), were used for experimental purposes with approval of the IICB Animal Ethical Committee and mice were handled according to their guidelines. Parasites and culture condition L. donovani, strain AG83 (MHOM/IN/1983/AG83) Mephenoxalone was originally isolated from an Indian kala-azar patient and maintained in Syrian golden hamsters by serial passage as described elsewhere [15]. Briefly, promastigotes were grown at 22°C in Medium 199 (pH 7.4) supplemented with 20% heat inactivated fetal bovine serum (FBS), 2 mM L-glutamine, 100 U/ml penicillin, 25 mM HEPES, 100 μg/ml streptomycin sulphate (all from Sigma-Aldrich, St. Louis, USA), and the parasites were subcultured in the same medium at an average density of 2 × 106 cells/ml at 22°C [15]. Preparation of leishmanial antigens LAg was prepared from L. donovani promastigotes as described earlier [15]. Briefly, stationary phase promastigotes, harvested after the third or fourth passage in liquid culture, were washed four times in cold 20 mM phosphate-buffered saline (PBS), pH 7.2, and resuspended at a concentration of 1.0 g cell pellet in 50 ml of cold 5 mM Tris-HCL buffer (pH 7.6).

9 0 0 2 8 0 0 Haemophilus 0 0 0 0 0 0 0 0 4 5 0 0 0 0 0 0 0 0 0 0

9 0.0 2.8 0.0 Haemophilus 0.0 0.0 0.0 0.0 4.5 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Peptoniphilus 0.0 2.9 6.3 0.0 0.0 38.6 7.1 11.5 50.4 0.0 9.1 0.0 Streptococcus 0.0 0.0 0.0 0.0 84.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Serratia 0.0 0.0 1.3 0.0 2.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 Percentages of each genera are indicated along with their location (A-L) based upon the map Selleckchem Inhibitor Library indicated in Figure 2B. Subject 3 A B C E G D F   E E E E E C C Peptoniphilus 32.1 62.5 49.4 54.2 13.9 44.0 9.6 Corynebacterium 10.7 3.8 2.8 0.0 15.6 0.0 13.1 Stenotrophomonas 14.2 0.0 0.0 0.0 0.0 0.0 0.0 Peptostreptococcus 7.1 6.2 7.7 6.1 6.5 0.0 1.1 Pseudomonas 17.8 7.5 17.1 0.0 21.3 12.0 11.8 Staphylococcus 7.1 2.5 2.3 0.0 31.0 20.0 41.0 Streptococcus 3.6 3.8 1.5 0.0 3.8 4.0 1.7

Acinetobacter 0.0 0.0 2.3 0.0 3.3 0.0 4.4 clostridia 0.0 7.5 3.8 5.5 1.6 8.0 1.5 Porphyromonas 0.0 1.3 0.0 23.7 1.6 0.0 4.3 Prevotella 0.0 0.0 3.6 0.0 0.0 4.0 0.0 Propionibacterium 0.0 0.0 0.0 0.0 0.0 8.0 0.0 Xanthomonas 0.0 0.0 0.0 0.0 0.0 12.0 0.0 Percentages of each genera are indicated along with their location (A-G) based upon the map indicated in Figure 2C. The location designations (edge or center) are also provided. Utilizing the new bTEFAP titanium technology a second topology evaluation was also conducted on 4 of the VLU patients. The new bTEFAP methods Acalabrutinib concentration utilize the new Titanium chemistry for pyrosequencing, which increases the read length of individual sequences

from an average of 250 bp to over 400 bp, utilize a single PCR step, and incorporate error reading polymerases. This new approach provides much better resolution at the individual species level and dramatically enhances our ability to characterize wound bacterial ecology. Four additional subjects were evaluated (See additional file 2). The results were similar to what we observed using the original bTEFAP method with the exception that we had more confidence in our ability to resolve certain populations at the species level. Subject 5 showed a high prevalence of Pseudomonas aeruginosa among the Exoribonuclease majority of the subsamples with notable populations of Burkholdaria spp (tentatively cenocepacia), an unknown Bacteroidales, and Clostridium spp (tentatively hathewayi). Subject 6 showed definite ubiquitous detection of Pseudomonas aeruginosa with notable populations of Streptococcus parasanguinis across the wound.

Quadruplet samples were run for each concentration of

Quadruplet samples were run for each concentration of Romidepsin cell line CH in three independent experiments. CH Treatment for a concentration- and Time-Dependent Study For a concentration- and time-dependent study, two sets of CH concentrations

(50 μg/mL and 150 μg/mL; 300 μg/mL and 600 μg/mL) were considered for treatment of MCF-7 cells for 24 hours. I found that 50 μg/mL CH did not show any significant induction of apoptosis whereas 600 μg/mL CH completely killed the cells. Hence, 150 μg/mL and 300 μg/mL concentrations of CH were used for further studies. MCF-7 cells were treated with either 150 μg/mL or 300 μg/mL CH for 24, 48 and 72 hours for the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. The cells were incubated with the same CHconcentrations for 24 and 48 hours for real-time quantitative PCR analysis. TUNEL Assay The DeadEnd® TUNEL assay kit (Promega, Madison, WI) was used for studying apoptosis in a time- and dose-dependent manner. The manufacturer’s instructions were followed with slight modifications. Briefly, MCF-7 cells Napabucasin in vivo (1.5 × 106 cells/well) were cultured in 6-well plates to study apoptosis in adherent cells. Cells were treated with 150 μg/mL and 300 μg/mL CH for 24, 48 and 72 hours. After the incubation period, the culture medium was aspirated

off, and the cell layers were trypsinized. The trypsinized cells were reattached on 0.01% polylysine-coated slides, fixed with 4% methanol-free formaldehyde solution, and stained according to the DeadEnd fluorometric TUNEL system protocol [16]. The stained cells were observed using a Carl-Zeiss (Axiovert) epifluorescence microscope using a triple band-pass filter. To determine the percentage of cells demonstrating apoptosis, 1000 cells were counted in each experiment [17]. Real-time quantitative PCR analysis The expression of apoptotic genes was analyzed

by reverse transcription-PCR (RT-PCR; Applied Biosystems 7500 Fast, Foster City, CA) using a real-time SYBR Green/ROX gene expression assay kit (QIAgen). The cDNA was directly prepared from cultured cells using a Fastlane® Cell cDNA kit (QIAGEN, Germany), and the mRNA levels of Caspase 3, Caspase 8, Caspase 9 and tp53 as well as the reference gene, GAPDH, were assayed using gene-specific SYBR Green-based QuantiTect® Ribonucleotide reductase Primer assays (QIAGEN, Germany). Quantitative real-time RT-PCR was performed in a reaction volume of 25 μL according to the manufacturer’s instructions. Briefly, 12.5 μL of master mix, 2.5 μL of primer assay (10×) and 10 μL of template cDNA (100 μg) were added to each well. After a brief centrifugation, the PCR plate was subjected to 35 cycles of the following conditions: (i) PCR activation at 95°C for 5 minutes, (ii) denaturation at 95°C for 5 seconds and (iii) annealing/extension at 60°C for 10 seconds. All samples and controls were run in triplicates on an ABI 7500 Fast Real-time PCR system.

The strategy differs from NOGG in that FRAX is always used with B

The strategy differs from NOGG in that FRAX is always used with BMD. Indeed, a BMD test is a prerequisite. Additionally, a fixed intervention threshold is used at all ages, whereas the NOGG strategy uses an age-dependent threshold. The rationale for a fixed threshold is based on the fracture probability at which intervention becomes cost-effective in the USA and the 20% threshold is, therefore, not relevant for any other country. Other assessment models As well as the FRAX tool, other fracture risk calculators are available online which include the Garvan fracture RGFP966 ic50 risk calculator and QFracture™ [69, 70]. Their comparative features are summarised in Table 9. The QFracture™ tool is based on

a UK prospective open cohort

study of routinely collected data from 357 general practices on over 2 million men and women aged 30–85 years (www.​qfracture.​org). Like the FRAX tool, it takes into account history of smoking, alcohol, corticosteroid use, parental history (of hip fracture or osteoporosis) and several secondary causes of osteoporosis. Unlike FRAX, it also includes a history of falls (yes/no only over an unspecified time frame) and excludes previous fracture history and BMD. It has been internally validated (i.e. from a stratum of the same population) and also externally validated in the UK [126]. Table 9 Comparative features of three fracture risk assessment algorithms   Dubbo/Garvan FK506 manufacturer Qfracture FRAX Externally validated Yes (a few countries) Yes (UK only) Yes Calibrated No Yes (UK only) Yes Applicability Unknown UK 45 countries Falls as an input variable Yesa Yes No BMD as an input variable Yes No Yes Prior fracture as an input variable Yesa No Yes Family history as an input variable No Yes Yes Output Incidence Incidence Probability Treatment responses assessed No No Yes aAnd number of falls/prior fractures The Garvan tool (www.​garvan.​org.​au) is based on data from participants enrolled in the Australian Dubbo Osteoporosis epidemiology study of approximately

2,500 men and next women age 60 years or more. It differs from FRAX by including a history of falls (categorised as 0, 1, 2 and >2 in the previous year) and the number of previous fragility fractures (categorised as 0, 1, 2 and >2), but does not include other FRAX variables. The output of the tool differs from FRAX in that it reports the risk of a larger number of fracture sites (additionally includes fractures of the distal femur, proximal tibia/fibula, distal tibia/fibula, patella, pelvis, ribs sternum, hands and feet excluding digits). As in the case of the QFracture, the Garvan tool captures fall risk. A fundamental difference between these risk models and FRAX is that the parameters of risk differ (incidence vs. probabilities) so that comparative data are not readily interpreted [127] (Fig. 10).

For this purpose, cells are grown in the light, either photoheter

For this purpose, cells are grown in the light, either photoheterotrophically in Tris acetate phosphate (TAP) or photoautotrophically in high salt minimal (HSM) medium (Harris 1989, 2009). For all the physiological analyses of Chlamydomonas and other algae, it is important to keep the cells of www.selleckchem.com/products/epz-6438.html precultures in the active growth phase. This means that as soon as the cultures have reached a cell density

of about 1 × 107 cells ml−1, an aliquot of this culture is used to inoculate fresh medium at a cell density of about 1–2 × 104 cells ml−1. For anaerobic adaptation of C. reinhardtii, a chlorophyll content of the pre-culture of 20–25 μg ml−1 has turned out to be optimal. The chlorophyll content of C. reinhardtii is determined by mixing 200 μl of cell suspension with 800 μl acetone, letting the chlorophylls extract for several hours in the refrigerator, spinning the cells down, and measuring the absorbance of the green supernatant against 80% acetone at λ = 652 nm (Arnon 1949). At a chlorophyll content of 20 μg ml−1, which is equivalent to about 6.5 × 106 cells ml−1 Tamoxifen datasheet in case of the C. reinhardtii wild type CC-124 (137c), the cells would have already reached the end of the exponential growth phase, but still divide. The pre-culture is then harvested by mild centrifugation (2 min at 3,500–5,000 g, room temperature) in Sarstedt (Sarstedt, Nümbrecht, Germany) or Falcon tubes and gently resuspended

in about 0.2 volumes of fresh medium very to reach a chlorophyll concentration of 100 μg ml−1. For reproducible and comparable results, both the respective pre-cultures and the concentrated cultures to be compared should have equivalent

chlorophyll contents. It is important for all the analyses of C. reinhardtii that the cells do not stand anywhere for more than 1 min, since they settle rapidly and establish microaerobic or even anaerobic conditions in the dense cell sediment. Accordingly, pellets of the algae should be resuspended rapidly. If the purpose of the experiments requires a real “0 h” sample, the concentrated cells may be incubated in a thin layer in Erlenmeyer flasks in the light to re-establish photosynthetic, thus O2-saturated conditions. For anaerobic adaptation, though the highly concentrated algal culture will not carry out appreciable photosynthesis due to self-shading, the tube should be further wrapped with aluminium foil to prevent the outer layers of the cell culture to be exposed to light. The algal suspension is then flushed by Ar or N2 at 18–25°C. For this purpose, a tube (about 5 mm in diameter) connected to the gas cylinder via a pressure regulator is introduced into the flask through a suitable hole in the lid, which should have a somewhat larger diameter than the tube to allow the gas to exhaust again. The tube is then dipped into the culture so that the gas flushes the cell suspension. It is important that the gas is of high purity, i.e., with no significant contaminations of O2.

This product was

purified and used as template for a seco

This product was

purified and used as template for a second PCR with the oligonucleotides Mal-C2Kpn and Ttrack2-U; the amplification product was named T2-U. A third PCR amplification product obtained with the primers RBS-C and Ttrack1-L, and pH3 DNA as the template, was purified and used as a template in a new PCR reaction with the primers RBS-C and Ttrack2-L. The amplification product was named T2-L. Finally, PCR products T2-U and T2-L were then mixed and used as the template for the last PCR. In this reaction, the CDK inhibitor primers Mal-C2Kpn and RBS-C were used, and the final PCR product was cloned into pDOP. Construction of repC hybrid genes Overlap extension PCR was also employed to obtain repC hybrid genes. RepC gene amplification products from pSymA were obtained using pDOP-CsA as the template, and the repC p42d products were obtained using pH3 as the template. Most of the hybrid genes described here required the overlap of two PCR products. The insert of plasmid

pDOP/C420-1209 was obtained using the primers C-SymA and AL-2Uc for the first PCR product and AL-2U and Mal-C2 for the second Selleck Ivacaftor product. The final PCR product was obtained with the external primers C-SymA and Mal-C2. The insert of plasmid pDOP/C1-420 was constructed with primers RBS-C and 1L-B2c and the primers 1L-B2 and K-SymAL for the first and second PCR products, respectively. These products were combined using the primers RBS-C and K-SymAL. The pDOP/C841-1209 insert was constructed with the primers C-SymA and BL-3Uc for the first PCR product and BL-3U and Mal-C2 for the second. These products were joined in a third PCR with the primers C-SymA and Mal-C2. The hybrid gene in pDOP/C1-990 was acquired with the primers RBS-C and Sal-CdL for the first PCR product and Sal-CdU and Mal-C2 for the second. These PCR products were integrated in a third PCR with the primers RBS-C and

Mal-C2. Similarly, the hybrid gene of pDOP/C1-990 was obtained with the primers RBS-C and Cd-1086 for the first amplification product. To obtain the second PCR product, the primers Cs-1087U and Mal-C2 were used, and both PCR products were fused with the primers RBS-C and Mal-C2. The inserts of two of the constructs, pDOP/C421-840 and pDOP/Cs421-840, required the fusion Unoprostone of three PCR products. The hybrid gene located in pDOP/C421-840 required the primers C-SymA and AL-2Uc for the first PCR product, the primers AL-2U and AL-2Uc for the second PCR product, and the primers 2L-CU and K-SymA for the third PCR product. The three PCR products were fused in the final PCR with the primers C-SymA and K-SymA. The hybrid gene present in pDOP/Cs421-840 was obtained using the primers RBS-C and 1L-B2c for the first PCR product, the primers 1L-B2 and B2-3Uc for the second PCR product, and the primers BL-2U and Mal-C2 for the third PCR product.

a: Control untreated cells;b: 0 008 μg/ml; c: 0 012 μg/ml, i e ,

a: Control untreated cells;b: 0.008 μg/ml; c: 0.012 μg/ml, i.e., the MIC dose; d: 0.04 μg/ml; e: 0.1 μg/ml; f: 0.5 μg/ml. The width of the dispersion of the fragments from the boundary of the nucleoid was quantified using an image analysis system; this measure is a simple and reliable quantitative parameter that reflects the level of CIP-induced DNA damage (Table 1). Differences were significant between the

doses tested from 0.012 learn more μg/ml, except between 0.012 μg/ml and 0.02 μg/ml, between 0.04 μg/ml and 0.08 μg/ml, and between 0.5 μg/ml and 1 μg/ml. Using the images obtained, the nucleoids were categorized into five classes of damage, as shown in Fig. 2 and Table 1: class 0: undamaged, dose of 0 to 0.008 μg/ml (Figs 1a and Selleck Venetoclax 1b); class I: low damage level, dose of 0.012 or 0.02 μg/ml (Fig. 1c); class II: intermediate level, dose of 0.04 or 0.08 μg/ml (Fig. 1d); class III: high level, dose of 0.1 μg/ml (Fig. 1e); and class IV: massive fragmentation, doses of 0.5 or 1 μg/ml or higher (Fig. 1f). This latter class of damage was practically undistinguishable from that shown by nucleoids with extensive DNA fragmentation always present spontaneously in cultures [15]. Classification into classes is standard practice in mutagenesis

studies and provides a perceptive description that is especially useful when heterogeneity in the DNA damage rank is evident between the different nucleoids, as observed in the DNA repair experiments. Table 1 Dose-response effect of CIP on TG1 E. coli chromosomal DNA analyzed with the Micro-Halomax® kit. Dose (μg/ml) Width of dispersion (μm) Class Range 0 –     0.003 – 0 0 0.006 –     0.008 –     0.012 1.3 ± 0.3 I ≤ 2.0 0.02 1.6 ± 0.3     0.04 2.5 ± 0.4 II 2.1 – 3.7 0.08 3.3 ± 0.4     0.1 5.1 ± 1.0 III 3.8 – 5.7 0.5 7.8 ± 1.4 IV ≥ 5.8 1 8.8 ± 1.6     The width of the halo of dispersion of DNA fragments is presented in μm (mean ± standard deviation). The extent of DNA damage was classified according to the width of the dispersion.

Leukotriene-A4 hydrolase Figure 2 Nucleoids from E. coli strain TG1 with progressively increased DNA fragmentation after incubation with increasing doses of CIP. 0: undamaged; I: low damage level; II: intermediate damage; III: high damage level; IV: massive fragmentation. Incubation time To determine the minimum incubation time needed to detect a DNA-breakage effect, the TG1 E. coli were collected from LB agar and exposed in liquid LB to 1 μg/ml CIP for 0, 5, 10, 15, 20, 30, and 40 min. The microgel preparation time before immersion in the lysing solution (8 min) must be added to these times because the antibiotic may enter the bacteria and act during this period. Detectable but subtle damage was apparent after 0 min (class I: diffusion width 1.7 ± 0.2 μm) (Fig. 3); this subtle damage appeared as nucleoids with some peripheral DNA fragments unlike in the untreated control cells.

Low-energy traumatic fractures (i e a fall from standing height)

Low-energy traumatic fractures (i.e. a fall from standing height) were regarded as osteoporotic fractures. Vertebral All spinal X-rays were taken according to local protocol; the same protocol was used at baseline and follow-up. Lateral radiographs of the spine were scored

according to the semi-quantitative method described by Genant et al. www.selleckchem.com/products/XL184.html [12]. Scoring was performed individually by two trained observers (MV and WL) and consensus both at baseline and follow-up was obtained in cases of discrepancies between both observers. Follow-up radiographs were scored blinded for the baseline image, and the results were subsequently compared to the baseline X-rays and scores to see if new vertebral fractures were detected. A fracture was scored as an incident vertebral fracture if it was not present at baseline or if there was a significant increase in loss of height (more than 20%) in a vertebra which was already fractures at baseline. Ethics The study protocol was approved by the local medical ethical committees of the three centres and all patients gave written informed consent. Statistical analysis Patients with incident fractures (vertebral or non-vertebral fractures) ZD1839 nmr were compared to those not having a new fracture with regard to demographic variables, clinical variables

and BMD using two-sided t tests for continuous variables and chi-square tests for counts. The incidence of patients with fractures was expressed per 100 patients/year with 95% confidence intervals (CI). Possible predictors of incident vertebral and non-vertebral fractures were subsequently examined in a multivariate logistic regression analysis. The criteria for entering from independent variables in the logistic regression analysis were a p value <0.2 in the univariate analysis and a supposed clinical relevance for the dependent variable. We were able

to build a prediction model with only significant covariates by using backward stepwise elimination of the least significant covariate. All statistical analyses were performed using SPSS (Chicago, IL, USA) version 15.0. Results Patient characteristics The clinical characteristics of the 102 patients included in this study are presented in Table 1. At baseline, the patients had a mean (SD) age of 61 (6) years with a median (range) disease duration of 17 (6–25) years, 83% of the patients had erosive disease and 65% patients were rheumatoid factor positive. Table 1 Characteristics of the 102 patients with RA included in the 5-year follow-up     Baseline Follow-up Age, years Mean (SD) 61 (6) na Disease duration, years Median (range) 17 (6–25) na IgM-RF positive (>25 U/ml) n (%) 67 (65) 67 (65) Joint erosions present, patients n (%) 85 (83) 85 (83) BMI, kg/m2 Mean (SD) 25.5 (5) 26.0 (5) HAQ Mean (SD) 1.48 (0.62) 1.59 (0.89) Corticosteroids Ever use n (%) 65 (64) na Use (during follow-up) n (%) na 58 (57)a Months used (during follow-up) Mean (SD) na 43.8 (25.4) ≥7.

1007/s003390051050CrossRef 32 Terrones M, Hsu WK, Kroto HW, Walt

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Nat Nanotechnol 2008, 3:210–215 CrossRef 40 Stampfer C, Molitor

Nat Nanotechnol 2008, 3:210–215.CrossRef 40. Stampfer C, Molitor F, Graf D, Ensslin K, Jungen A, Hierold C, Ensslin K: Raman imaging of doping domains in graphene on SiO(2). Appl Phys Lett 2007, 91:241907.CrossRef Competing interests The authors declare

that they have no competing interests. Authors’ contributions C-H and B-JL carried on the experimental parts: the acquisition of data and analysis and interpretation of data. C-H also had been involved in drafting the manuscript. H-YL and C-HH analyzed and interpreted the data. They also had been involved in revising the manuscript. F-YS and W-HW (Institute of Atomic and Molecular Sciences, Academia Sinica) prepared the samples, suspended graphene using by micromechanical Kinase Inhibitor Library in vitro method, and captured the OM and AFM images. C-YL have made substantial contributions click here to the conception and design of the study and revising it critically for important intellectual content. H-CC, the corresponding author, had made substantial contributions to the conception and design of the study and had been involved in drafting the manuscript and revised it critically for important intellectual content. All authors read and approved the final manuscript.”

Scanning tunneling microscopy (STM) [1] and atomic force microscopy (AFM) [2] have revolutionized surface sciences by enabling the study of surface topography and other surface 3-mercaptopyruvate sulfurtransferase properties at the angstrom-to-micrometer scale. The three major functions of AFM include imaging, spectroscopy (i.e., force-distance curve), and manipulation (nanolithography).

AFM techniques employ a very sharp tip as a probe to scan and image surfaces. Spectroscopic information is acquired through forces generated between the tip and the sample when the probe is brought into proximity with the sample surface, according to Hooke’s law. Xie et al. [3] classified nanolithographic techniques into two groups: force-assisted and bias-assisted nanolithography. In AFM, the interactive force between the tip of the probe and the sample surface is determined according to the deflection of a microfabricated cantilever with the tip positioned at the free end. Modifying the probe enables researchers to explore a range of surface characteristics. AFM probes with individual microparticles or nanoparticles attached to the cantilever/tip have been widely used to measure surface forces in AFM and near-field scanning optical microscopy (NSOM) [4] as the geometry and composition of the particle can be well controlled. Ducker et al. [5, 6] were pioneers in the attachment of microspheres to a tipless AFM cantilever with resin. Their colloidal probe technique employed a laser-pulled micropipette attached to an optical microscope. Mak et al.